Tubulin inhibitors

ABSTRACT

Compounds of general formula (I), (II), (III) and (V) are described for use in modulating microtubule polymerisation and in the treatment of associated disease states. Use of compounds (I), (III) and (V) in the treatment of kinase-associated disease states is also described. Further described are novel compounds of formula (II), (III) and (V).

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. Ser. No. 10/581,534 with aninternational filing date of 3 Dec. 2004, and now allowed, which claimsbenefit under 35 U.S.C. §119(e) to International application No.PCT/AU2004/001689 filed 3 Dec. 2004, which claims benefit of AustralianApplication No. 2003906680 filed 3 Dec. 2003. The contents of theseapplications are incorporated herein by reference.

FIELD OF THE INVENTION

The present invention relates to methods of suppressing the growth ofcancers and other proliferative diseases by the administration ofcompounds that act by binding to tubulin and to novel heterocycliccompounds suitable for such administration.

BACKGROUND OF THE INVENTION

There are many human and veterinary diseases that stem from processes ofuncontrolled or abnormal cellular proliferation. Most important amongthese diseases is cancer, the generic name given to a broad range ofcellular malignancies characterized by unregulated growth and lack ofdifferentiation. Psoriasis is another disease that is characterized byuncontrolled or abnormal cellular proliferation. Psoriasis is a commonchronic skin disease characterized by the presence of dry scales andplaques. The disease results from hyperproliferation of the epidermisand incomplete differentiation of keratinocytes. Psoriasis ofteninvolves the scalp, elbows, knees, back, buttocks, nails, eyebrows, andgenital regions, and may range in severity from mild to extremelydebilitating, resulting in psoriatic arthritis, pustular psoriasis, andexfoliative psoriatic dermatitis. There is, at present, no generaltherapeutic cure that exists for psoriasis. Whilst milder cases areoften treated with topical cortico-steroids, more severe cases may betreated with antiproliferative agents, such as the antimetabolitemethotrexate, the DNA synthesis inhibitor hydroxyurea, and themicrotubule disrupter colchicine.

Other diseases associated with an abnormally high level of cellularproliferation include restenosis, where vascular smooth muscle cells areinvolved; inflammatory disease states, where endothelial cells,inflammatory cells and glomerular cells are involved; myocardialinfarction, where heart muscle cells are involved; glomerular nephritis,where kidney cells are involved; transplant rejection, where endothelialcells are involved; and infectious diseases such as HIV infection andmalaria, where certain immune cells and/or other infected cells areinvolved.

Inhibition of cellular proliferation can be brought about by severalmechanisms, including; alkylating agents; topoisomerase inhibitors;nucleotide analogues; antibiotics; hormone antagonists; and nucleic aciddamaging agents; inter alia. One pharmacologically important mechanismof inhibiting cellular proliferation is by means of binding tubulin.Tubulin is an asymmetric dimer composed of alpha and beta subunits, thatpolymerizes to form structural components of the cytoskeleton calledmicrotubules. Microtubules must be highly dynamic in order to carry outmany of their functions. At certain stages of the cell cycle, or inparticular cell types or organelles, stable microtubules are required,such as for transport within axons or for ciliary and flagellarmovement. Micro-tubules assemble during the G2 phase of the cell cycle,and participate in the formation of the mitotic spindle whichfacilitates the segregation of sister chromatids during the process ofcell division. The essential role of microtubules in cell division andthe ability of drugs that interact with tubulin to interfere with thecell cycle have made tubulin a successful target for applications thatinclude anti-cancer drugs, fungicides, and herbicides. Typical tubulinligands such as colchicine, paclitaxel, the Vinca alkaloids such asvinblastine, the epothilones, the halicondrins, benomyl and mebendazoledirectly inhibit cell division by binding to tubulin which leads to thearrest of the cell cycle at the G2/M boundary of mitosis. This mechanismis the basis of the therapeutic value of compounds of this type, such astreating gout with colchicine, restenosis with paclitaxel, cancer withpaclitaxel, vinblastine, epothilones and halichondrins, and fungalinfections with benomyl and malaria and helminths with mebendazole.

Interfering with microtubule dynamics or stability can inhibit celldivision in several ways. Both stablilizing microtubules or inhibitingtheir polymerization will prevent the cytoskeleton restructuring that isrequired at several points in the cell cycle and lead to an arrest ofthe cell's progression from one stage in the cell cycle to the next.Three main classes of tubulin-binding drugs, namely colchicineanalogues, Vinca alkaloids, and the taxanes, have been identified, eachof which possesses a specific binding site on the β-tubulin molecule.Paclitaxel (Taxol™) and related taxanes represent a class of drugs thatstabilize microtubules, a process that ultimately leads to the“freezing” of the microtubule structures so that they cannot berestructured (Jordan M. A. and Wilson L., 1998). Subsequent arrest atmitosis induces the apoptotic mechanism to cause cell death. A number ofcolchicine analogues, as well as several other compounds that bind tothe same site on β-tubulin as colchicine disrupt tubulin polymerizationand disrupt microtubular formation. Vinblastine and several othervinca-related drugs bind to a site that is distinct from the colchicinesite. Compounds that bind at the Vinca-site prevent microtubuleformation and destabilize microtubules (Jordan et al, 1986; Rai andWolff (1996). This invention is therefore directed to compounds thatpotentially modulate microtubule dynamics by binding to tubulin.

Accordingly, the present invention aims to provide compounds which aredirectly or indirectly toxic to actively dividing cells and are usefulin the treatment of cancer, viral and bacterial infections, vascularrestenosis, inflammatory diseases, autoimmune diseases, or psoriasis.The present invention is also directed to therapeutic compositions fortreating said conditions. Further aspects of the invention are toprovide methods for killing actively proliferating cells, such ascancerous, bacterial, or epithelial cells, and treating all types ofcancers, infections, inflammatory, and generally proliferativeconditions. A further aspect relates to provide methods for treatingother medical conditions characterized by the presence of rapidlyproliferating cells, such as psoriasis and other skin disorders.

In one embodiment, the method of the invention is used in the treatmentof sarcomas, carcinomas and/or leukemias. Exemplary disorders for whichthe subject method can be used alone or as part of a treatment regimeninclude: fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma,osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma,lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma,Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon carcinoma,pancreatic cancer, breast cancer, ovarian cancer, prostate cancer,squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweatgland carcinoma, sebaceous gland carcinoma, papillary carcinoma,papillary adenocarcinomas, cystadenocarcinoma, medullary carcinoma,bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile ductcarcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilms' tumor,cervical cancer, testicular tumor, lung carcinoma, small cell lungcarcinoma, bladder carcinoma, epithelial carcinoma, glioma, astrocytoma,medulloblastoma, craniopharyngioma, ependymoma, pinealoma,hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma,melanoma, neuroblastoma, and retinoblastoma.

In certain embodiments, the method of the invention is to be used totreat disorders such as carcinomas forming from tissue of the breast,prostate, kidney, bladder or colon.

In other embodiments, the method of the invention is used to treathyperplastic or neoplastic disorders arising in adipose tissue, such asadipose cell tumors, e.g., lipomas, fibrolipomas, lipoblastomas,lipomatosis, hibemomas, hemangiomas and/or liposarcomas.

In still other embodiments, infectious and parasitic agents (e.g.bacteria, trypanosomes, fungi, etc.) can also be controlled using thesubject compositions and compounds.

SUMMARY OF THE INVENTION

The present inventors have found that a group of compounds based upon asubstituted heterocyclic scaffold inhibit the growth and proliferationof cancer cells. The present inventors have further shown that thesecompounds can bind to tubulin. Such compounds would also be useful inthe treatment of other hyperproliferation related disorders.

Accordingly, in a first aspect the present invention provides a methodof modulating microtubule polymerisation in a subject, the methodcomprising administering a therapeutically effective amount of at leastone compound of the general formula

or pharmaceutically acceptable prodrugs, salts, hydrates, solvates,crystal forms or diastereomers thereof, wherein:

-   -   R1 is H, C₁₋₄ alkyl;    -   Q is a bond, or C₁₋₄ alkyl;    -   A is aryl, hetaryl optionally substituted with 0-3 substituents        independently chosen from halogen, C₁₋₄ alkyl, CH₂F, CHF₂, CF₃,        CN, aryl, hetaryl, OCF₃, OC₁₋₄alkyl, OC₂₋₅alkylNR4R5, Oaryl,        Ohetaryl, CO₂R4, CONR4R5, nitro, NR4R5, C₁₋₄ alkylNR4R5,        NR6C₁₋₄alkylNR4R5, NR4COR5, NR6CONR4R5, NR4SO₂R5;        -   R4, R5 are each independently H, C₁₋₄ alkyl, C₁₋₄ alkyl            cycloalkyl, C₁₋₄ alkyl cyclohetalkyl, aryl, hetaryl,            C₁₋₄alkyl aryl, C₁₋₄ alkyl hetaryl, or may be joined to form            an optionally substituted 3-8 membered ring optionally            containing an atom selected from O, S, NR7;        -   R6 is selected from H, C₁₋₄ alkyl;            -   R7 is selected from H, C₁₋₄ alkyl, aryl, hetaryl, C₁₋₄                alkyl aryl, C₁₋₄ alkyl hetaryl;    -   R2 is 0-2 substituents independently selected from halogen,        C₁₋₄alkyl, OH, OC₁₋₄alkyl, CH₂F, CHF₂, CF₃, OCF₃, CN,        C₁₋₄alkylNR8R9, OC₁₋₄alkylNR8R9, CO₂R8, CONR8R9, NR8R9, NR8COR9,        NR10CONR8R9, NR8SO₂R9;        -   R8, R9 are each independently H, C₁₋₄ alkyl, C₁₋₄ alkyl            cycloalkyl, C₁₋₄ alkyl cyclohetalkyl, aryl, hetaryl, C₁₋₄            alkyl aryl, C₁₋₄ alkyl hetaryl, or may be joined to form an            optionally substituted 3-8 membered ring optionally            containing an atom selected from O, S, NR11;        -   R10 is selected from H, C₁₋₄ alkyl, aryl or hetaryl;            -   R11 is selected from H, C₁₋₄ alkyl, aryl, hetaryl, C₁₋₄                alkyl aryl, C₁₋₄ alkyl hetaryl;    -   Y is halogen, OH, NR12R13, NR14COR12, NR14CONR12R13, N14SO₂R13;        -   R12 and R13 are each independently H, CH₂F, CHF₂, CF₃, CN,            C₁₋₄ alkyl optionally substituted with OH, OC₁₋₄alkyl or            NR15R16, cycloalkyl; cyclohetalkyl, C₁₋₄ alkyl cycloalkyl,            C₁₋₄ alkyl cyclohetalkyl, or may be joined to form an            optionally substituted 3-6 membered ring optionally            containing an atom selected from O, S, NR14            -   R14, R15 and R16 are each independently selected from H,                C₁₋₄ alkyl;    -   n=0-4;    -   W is selected from H, C₁₋₄alkyl, C₂₋₆alkenyl; where C₁₋₄alkyl or        C₂₋₆alkenyl may be optionally substituted with C₁₋₄alkyl, OH,        OC₁₋₄alkyl, NR15R16;        -   R15, and R16 are each independently H, C₁₋₄ alkyl, C₁₋₄            alkyl cycloalkyl, C₁₋₄ alkyl cyclohetalkyl, or may be joined            to form an optionally substituted 3-8 membered ring            optionally containing an atom selected from O, S, NR17;            -   R17 is selected from H, C₁₋₄ alkyl.

In a second aspect the present invention provides a compound of thegeneral formula

or pharmaceutically acceptable prodrugs, salts, hydrates, solvates,crystal forms or diastereomers thereof, wherein:

-   -   R1 is H, C₁₋₄ alkyl;    -   Q is a bond, or C₁₋₄ alkyl;    -   A is aryl, hetaryl optionally substituted with 0-3 substituents        independently chosen from halogen, C₁₋₄ alkyl, CH₂F, CHF₂, CF₃,        CN, aryl, hetaryl, OCF₃, OC₁₋₄alkyl, OC₂₋₅alkylNR4R5, Oaryl,        Ohetaryl, CO₂R4, CONR4R5, nitro, NR4R5, C₁₋₄ alkylNR4R5,        NR6C₁₋₄alkylNR4R5, NR4COR5, NR6CONR4R5, NR4SO₂R5;        -   R4, R5 are each independently H, C₁₋₄ alkyl, C₁₋₄ alkyl            cycloalkyl, C₁₋₄ alkyl cyclohetalkyl, aryl, hetaryl,            C₁₋₄alkyl aryl, C₁₋₄ alkyl hetaryl, or may be joined to form            an optionally substituted 3-8 membered ring optionally            containing an atom selected from O, S, NR7;        -   R6 is selected from H, C₁₋₄ alkyl;            -   R7 is selected from H, C₁₋₄ alkyl, aryl, hetaryl, C₁₋₄                alkyl aryl, C₁₋₄ alkyl hetaryl;    -   R2 is 0-2 substituents independently selected from C₁₋₄alkyl and        OC₁₋₄alkyl;    -   Y is CH₂OH, OC₁₋₄alkylOH, OC₁₋₄alkylR12, OC₁₋₄alkylNR12NR13,        C(O)R12, CH₂R12, COOR12, CONR12R13, OCONR12R13, CH₂NR12R13,        NHCOR12, NHCONR12R13,        -   R12 and R13 are each independently H, C₁₋₂ alkyl,            (CH₂)₃NEt₂, (CH₂)₂NMe₂, (CH₂)₅NH₂, (CH₂)₂OH,

-   -   W is selected from H, C₁₋₄alkyl, C₂₋₆alkenyl; where C₁₋₄alkyl or        C₂₋₆alkenyl may be optionally substituted with C₁₋₄alkyl, OH,        OC₁₋₄alkyl, NR15R16;        -   R15, and R16 are each independently H, C₁₋₄ alkyl, C₁₋₄            alkyl cycloalkyl, C₁₋₄ alkyl cyclohetalkyl, or may be joined            to form an optionally substituted 3-8 membered ring            optionally containing an atom selected from O, S, NR17            -   R17 is selected from H, C₁₋₄ alkyl;    -   wherein when Y is CH₂R12 then R12 is not H, C₁₋₂alkyl.

In a third aspect, the present invention provides a compositioncomprising a carrier and at least one compound of the second aspect.

In a fourth aspect the present invention provides a method of treating ahyperproliferation-related disorder in a subject, the method comprisingadministering a therapeutically effective amount of at least onecompound of the second aspect or a composition of the third aspect.

In a fifth aspect the present invention provides a compound of thegeneral formula

or pharmaceutically acceptable prodrugs, salts, hydrates, solvates,crystal forms or diastereomers thereof, wherein:

-   -   X₁, X₂, X₃, X₄ are selected from the following:    -   (i) X₁ and X₂ are N and X₃ and X₄ are C independently        substituted with Y;    -   (ii) X₁ and X₄ are N and X₂ and X₃ are C independently        substituted with Y;    -   (iii) X₁ and X₃ are N and X₂ and X₄ are C independently        substituted with Y;    -   (iv) X₂ and X₄ are N and X₁ and X₃ are C independently        substituted with Y;    -   (v) X₁ is N and X₂, X₃, and X₄ are C independently substituted        with Y;    -   (vi) X₃ is N and X₁, X₂, and X₄ are C independently substituted        with Y;    -   (vii) X₄ is N and X₁, X₂, and X₃ are C independently substituted        with Y;    -   (viii) X₂ is N and X₁, X₃, and X₄ are C independently        substituted with Y; and    -   (ix) X₁, X₂ and X₃ are N and X₄ is C substituted with Y;    -   R1 is H, C₁₋₆alkyl, C₁₋₆ alkylNR5R6, C₁₋₆alkylNR5COR6,        C₁₋₆alkylNR5SO₂R6, C₁₋₆alkylCO₂R5, C₁₋₆alkylCONR5R6;        -   R5 and R6 are each independently H, C₁₋₄alkyl, aryl,            hetaryl, C₁₋₄alkylaryl, C₁₋₄alkylhetaryl or may be joined to            form an optionally substituted 3-8 membered ring optionally            containing an atom selected from O, S, NR7;            -   R7 is selected from H, C₁₋₄alkyl;    -   R2 is selected from C₁₋₆alkylOH, OC₂₋₆alkylOH, C₁₋₆alkylNR8R9,        OC₂₋₆alkylNR8R9, C₁₋₆alkylNR8COR9, OC₂₋₆alkylNR8COR9,        C₁₋₆alkylhetaryl, OC₂₋₆alkylhetaryl, OCONR8R9, NR8COOR9,        NR10CONR8R9, CONR8R9, NR8COR12;        -   R8, R9 are each independently H, C₁₋₄alkyl,            C₁₋₄alkylNR11R13, hetaryl, cyclohetalkyl, or may be joined            to form an optionally substituted 3-8 membered ring            optionally containing an atom selected from O, S, NR14;        -   R12 is C₂₋₄alkyl, C₁₋₄alkylNR11R13, hetaryl, cyclohetalkyl;            -   R11, R13 are each independently H, C₁₋₄alkyl, or may be                joined to form an optionally substituted 3-8 membered                ring optionally containing an atom selected from O, S,                NR14;                -   R14 is selected from H, C₁₋₄alkyl;        -   R10 is H, C₁₋₄ alkyl;    -   R3 and R4 are each independently H, halogen, C₁₋₄alkyl, OH,        OC₁₋₄alkyl, CF₃, OCF₃;    -   Q is a bond, or C₁₋₄ alkyl;    -   W is selected from H, C₁₋₄alkyl, C₂₋₆alkenyl; where C₁₋₄alkyl or        C₂₋₆alkenyl may be optionally substituted with C₁₋₄alkyl, OH,        OC₁₋₄alkyl, NR15R16;        -   R15, and R16 are each independently H, C₁₋₄alkyl cycloalkyl,            C₁₋₄alkyl cyclohetalkyl, aryl, hetaryl, or may be joined to            form an optionally substituted 3-8 membered ring optionally            containing an atom selected from O, S, NR17;            -   R17 is selected from H, C₁₋₄alkyl;    -   A is aryl, hetaryl optionally substituted with 0-3 substituents        independently chosen from halogen, C₁₋₄ alkyl, CF₃, aryl,        hetaryl, OCF₃, OC₁₋₄alkyl, OC₂₋₅alkylNR18R19, Oaryl, Ohetaryl,        CO₂R18, CONR18R19, NR18R19, C₁₋₄ alkylNR18R19,        NR2OC₁₋₄alkylNR18R19, NR18COR19, NR20CONR18R19, NR18SO₂R19;        -   R18, R19 are each independently H, C₁₋₄ alkyl, C₁₋₄ alkyl            cyclohetalkyl, aryl, hetaryl, C₁₋₄alkyl aryl, C₁₋₄ alkyl            hetaryl, or may be joined to form an optionally substituted            3-8 membered ring optionally containing an atom selected            from O, S, NR21;            -   R21 is selected from H, C₁₋₄alkyl;        -   R20 is selected from H, C₁₋₄alkyl;    -   Y is selected from H, C₁₋₄alkyl, OH, NR22R23;        -   R22, R23 are each independently H, C₁₋₄alkyl.

In a sixth aspect, the present invention provides a compositioncomprising a carrier and one or more compounds of the fifth aspect.

In a seventh aspect, the present invention provides a method oftreatment of a hyperproliferation-related disorder in a subject, themethod comprising administering a therapeutically effective amount of atleast one of the compounds of the fifth aspect or a compositionaccording to the sixth aspect.

In an eighth aspect, the present invention consists in a method oftreating a protein-kinase related disorder in a subject, the methodcomprising administering a therapeutically effective amount of at leastone of the compounds of the fifth aspect or a composition according tothe sixth aspect.

In a ninth aspect, the present invention provides a compound of thegeneral formula

or pharmaceutically acceptable prodrugs, salts, hydrates, solvates,crystal forms or diastereomers thereof, wherein:

-   -   X₁, X₂, X₃, X₄ are selected from the following:    -   (i) X₁ and X₂ are N and X₃ and X₄ are C independently        substituted with Y;    -   (ii) X₁ and X₄ are N and X₂ and X₃ are C independently        substituted with Y;    -   (iii) X₂ and X₄ are N and X₁ and X₃ are C independently        substituted with Y;    -   (iv) X₁ is N and X₂, X₃, and X₄ are C independently substituted        with Y;    -   (v) X₃ is N and X₁, X₂, and X₄ are C independently substituted        with Y;    -   (vi) X₄ is N and X₁, X₂, and X₃ are C independently substituted        with Y;    -   (vii) X₂ is N and X₁, X₃, and X₄ are C independently substituted        with Y; and    -   (viii) X₁, X₂ and X₃ are N and X₄ is C substituted with Y;    -   R1 is H, C₁₋₆alkyl, C₁₋₆ alkylNR5R6, C₁₋₆alkylNR5COR6,        C₁₋₆alkylNR5SO₂R6, C₁₋₆alkylCO₂R5, C₁₋₆alkylCONR5R6;        -   R5 and R6 are each independently H, C₁₋₄alkyl, aryl,            hetaryl, C₁₋₄alkylaryl, C₁₋₄alkylhetaryl or may be joined to            form an optionally substituted 3-8 membered ring optionally            containing an atom selected from O, S, NR7;            -   R7 is selected from H, C₁₋₄ alkyl;    -   R2 is selected from OH, OC₁₋₆alkyl, C₁₋₆alkylOH, OC₂₋₆alkylOH,        C₁₋₆alkylNR8R9, OC₂₋₆alkylNR8R9, C₁₋₆alkylNR8COR9,        OC₂₋₆alkylNR8COR9, C₁₋₆alkylhetaryl, OC₂₋₆alkylhetaryl,        OCONR8R9, NR8COOR9, NR10CONR8R9, CONR8R9, NR8COR12;        -   R8, R9 are each independently H, C₁₋₄alkyl,            C₁₋₄alkylNR11R13, hetaryl, cyclohetalkyl, or may be joined            to form an optionally substituted 3-8 membered ring            optionally containing an atom selected from O, S, NR14;        -   R12 is C₂₋₄alkyl, C₁₋₄alkylNR11R13, hetaryl, cyclohetalkyl;            -   R11, R13 are each independently H, C₁₋₄alkyl, or may be                joined to form an optionally substituted 3-8 membered                ring optionally containing an atom selected from O, S,                NR14;                -   R14 is selected from H, C₁₋₄alkyl;        -   R10 is H, C₁₋₄ alkyl;    -   R3 and R4 are each independently H, halogen, C₁₋₄alkyl, OH,        OC₁₋₄alkyl, CF₃, OCF₃;    -   Q is a bond, or C₁₋₄alkyl;    -   W is selected from H, C₁₋₄alkyl, C₂₋₆alkenyl; where C₁₋₄alkyl or        C₂₋₆alkenyl may be optionally substituted with C₁₋₄alkyl, OH,        OC₁₋₄alkyl, NR15R16;        -   R15, and R16 are each independently H, C₁₋₄alkyl, C₁₋₄alkyl            cycloalkyl, C₁₋₄alkyl cyclohetalkyl, aryl, hetaryl, or may            be joined to form an optionally substituted 3-8 membered            ring optionally containing an atom selected from O, S, NR17;            -   R17 is selected from H, C₁₋₄alkyl;    -   A is aryl, hetaryl optionally substituted with 0-3 substituents        independently chosen from halogen, C₁₋₄ alkyl, CF₃, aryl,        hetaryl, OCF₃, OC₁₋₄alkyl, OC₂₋₅alkylNR18R19, Oaryl, Ohetaryl,        CO₂R18, CONR18R19, NR18R19, C₁₋₄ alkylNR18R19,        NR2OC₁₋₄alkylNR18R19, NR18COR19, NR20CONR18R19, NR18SO₂R19;        -   R18, R19 are each independently H, C₁₋₄ alkyl, C₁₋₄ alkyl            cyclohetalkyl, aryl, hetaryl, C₁₋₄alkyl aryl, C₁₋₄ alkyl            hetaryl, or may be joined to form an optionally substituted            3-8 membered ring optionally containing an atom selected            from O, S, NR21;            -   R21 is selected from H, C₁₋₄ alkyl;        -   R20 is selected from H, C₁₋₄ alkyl;    -   Y is selected from H, C₁₋₄alkyl, OH, NR22R23;        -   R22, R23 are each independently H, C₁₋₄ alkyl.

In a tenth aspect, the present invention provides a compositioncomprising a carrier and one or more compounds of the ninth aspect.

In an eleventh aspect, the present invention provides a method oftreatment of a hyperproliferation-related disorder in a subject, themethod comprising administering a therapeutically effective amount of atleast one of the compounds of the ninth aspect or a compositionaccording to the tenth aspect.

In a twelfth aspect, the present invention consists in a method oftreating a protein-kinase related disorder in a subject, the methodcomprising administering a therapeutically effective amount of at leastone of the compounds of the ninth aspect or a composition according tothe tenth aspect.

DETAILED DESCRIPTION OF THE INVENTION

In a first aspect the present invention provides a method of modulatingmicrotubule polymerisation in a subject, the method comprisingadministering a therapeutically effective amount of at least onecompound of the general formula

or pharmaceutically acceptable salts, hydrates, solvates, crystal formsor diastereomers thereof, wherein:

-   -   R1 is H, C₁₋₄ alkyl;    -   Q is a bond, or C₁₋₄ alkyl;    -   A is aryl, hetaryl optionally substituted with 0-3 substituents        independently chosen from halogen, C₁₋₄ alkyl, CH₂F, CHF₂, CF₃,        CN, aryl, hetaryl, OCF₃, OC₁₋₄alkyl, OC₂₋₅alkylNR4R5, Oaryl,        Ohetaryl, CO₂R4, CONR4R5, nitro, NR4R5, C₁₋₄ alkylNR4R5,        NR6C₁₋₄alkylNR4R5, NR4COR5, NR6CONR4R5, NR4SO₂R5; and R4, R5 are        each independently H, C₁₋₄ alkyl, C₁₋₄ alkyl cycloalkyl, C₁₋₄        alkyl cyclohetalkyl, aryl, hetaryl, C₁₋₄alkyl aryl, C₁₋₄ alkyl        hetaryl, or may be joined to form an optionally substituted 3-8        membered ring optionally containing an atom selected from O, S,        NR7; and R6 is selected from H, C₁₋₄ alkyl; and R7 is selected        from H, C₁₋₄ alkyl, aryl, hetaryl, C₁₋₄ alkyl aryl, C₁₋₄ alkyl        hetaryl;    -   R2 is 0-2 substituents independently selected from halogen,        C₁₋₄alkyl, OH, OC₁₋₄alkyl, CH₂F, CHF₂, CF₃, OCF₃, CN,        C₁₋₄alkylNR8R9, OC₁₋₄alkylNR8R9, CO₂R8, CONR8R9, NR8R9, NR8COR9,        NR10CONR8R9, NR8SO₂R9; and R8, R9 are each independently H, C₁₋₄        alkyl, C₁₋₄ alkyl cycloalkyl, C₁₋₄ alkyl cyclohetalkyl, aryl,        hetaryl, C₁₋₄ alkyl aryl, C₁₋₄ alkyl hetaryl, or may be joined        to form an optionally substituted 3-8 membered ring optionally        containing an atom selected from O, S, NR11; and R10 is selected        from H, C₁₋₄ alkyl, aryl or hetaryl; and R11 is selected from H,        C₁₋₄ alkyl, aryl, hetaryl, C₁₋₄ alkyl aryl, C₁₋₄ alkyl hetaryl;    -   Y is halogen; OH; NR12R13; NR14COR12; NR14CONR12R13; N14SO₂R13;        and R12 and R13 are each independently H; CH₂F; CHF₂; CF₃; CN;        C₁₋₄ alkyl optionally substituted with OH, OC₁₋₄alkyl or        NR15R16; cycloalkyl; cyclohetalkyl; C₁₋₄ alkyl cycloalkyl; C₁₋₄        alkyl cyclohetalkyl; or may be joined to form an optionally        substituted 3-6 membered ring optionally containing an atom        selected from O, S, NR14 and R14, R15 and R16 are each        independently selected from H, C₁₋₄ alkyl;    -   n=0-4;    -   W is selected from H, C₁₋₄alkyl, C₂₋₆alkenyl; where C₁₋₄alkyl or        C₂₋₆alkenyl may be optionally substituted with C₁₋₄alkyl, OH,        OC₁₋₄alkyl, NR15R16; and R15, and R16 are each independently H,        C₁₋₄ alkyl, C₁₋₄ alkyl cycloalkyl, C₁₋₄ alkyl cyclohetalkyl, or        may be joined to form an optionally substituted 3-8 membered        ring optionally containing an atom selected from O, S, NR17 and        R17 is selected from H, C₁₋₄ alkyl.

Preferably, the method is used in the treatment of ahyperproliferation-related disorder.

Preferably, the compound is selected from the compounds set out in Table2.

In a second aspect the present invention provides a compound of thegeneral formula

or pharmaceutically acceptable prodrugs, salts, hydrates, solvates,crystal forms or diastereomers thereof, wherein:

-   -   R1 is H, C₁₋₄ alkyl;    -   Q is a bond, or C₁₋₄ alkyl;    -   A is aryl, hetaryl optionally substituted with 0-3 substituents        independently chosen from halogen, C₁₋₄ alkyl, CH₂F, CHF₂, CF₃,        CN, aryl, hetaryl, OCF₃, OC₁₋₄alkyl, OC₂₋₅alkylNR4R5, Oaryl,        Ohetaryl, CO₂R4, CONR4R5, nitro, NR4R5, C₁₋₄ alkylNR4R5,        NR6C₁₋₄alkylNR4R5, NR4COR5, NR6CONR4R5, NR4SO₂R5; and R4, R5 are        each independently H, C₁₋₄ alkyl, C₁₋₄ alkyl cycloalkyl, C₁₋₄        alkyl cyclohetalkyl, aryl, hetaryl, C₁₋₄alkyl aryl, C₁₋₄ alkyl        hetaryl, or may be joined to form an optionally substituted 3-8        membered ring optionally containing an atom selected from O, S,        NR7; and R6 is selected from H, C₁₋₄ alkyl; and R7 is selected        from H, C₁₋₄ alkyl, aryl, hetaryl, C₁₋₄ alkyl aryl, C₁₋₄ alkyl        hetaryl;    -   R2 is 0-2 substituents independently selected from C₁₋₄alkyl and        OC₁₋₄alkyl;    -   Y is CH₂OH, OC₁₋₄alkylR12, OC₁₋₄alkylNR12NR13, C(O)R12, CH₂R12,        COOR12, CONR12R13, OCONR12R13, CH₂NR12R13, NHCOR12, NHCONR12R13,        and R12 and R13 are each independently H, C₁₋₂ alkyl,        (CH₂)₃NEt₂, (CH₂)₂NMe₂, (CH₂)₅NH₂, (CH₂)₂OH,

-   -   W is selected from H, C₁₋₄alkyl, C₂₋₆alkenyl; where C₁₋₄alkyl or        C₂₋₆alkenyl may be optionally substituted with C₁₋₄alkyl, OH,        OC₁₋₄alkyl, NR15R16; and R15, and R16 are each independently H,        C₁₋₄ alkyl, C₁₋₄ alkyl cycloalkyl, C₁₋₄ alkyl cyclohetalkyl, or        may be joined to form an optionally substituted 3-8 membered        ring optionally containing an atom selected from O, S, NR17 and        R17 is selected from H, C₁₋₄ alkyl;    -   wherein when Y is CH₂R12 then R12 is not H, C₁₋₂alkyl.

In a third aspect, the present invention comprises a compositioncomprising a carrier and at least one compound of the second aspect.

In a fourth aspect the present invention comprises a method of treatinga hyperproliferation-related disorder in a subject, the methodcomprising administering a therapeutically effective amount of at leastone compound of the second aspect or a composition of the third aspect.

Preferably, the hyperproliferation-related disorder is treatable by themodulation of microtubule polymerisation.

In a fifth aspect the present invention provides a compound of thegeneral formula

or pharmaceutically acceptable prodrugs, salts, hydrates, solvates,crystal forms or diastereomers thereof, wherein:

-   -   X₁, X₂, X₃, X₄ are selected from the following:    -   (i) X₁ and X₂ are N and X₃ and X₄ are C independently        substituted with Y;    -   (ii) X₁ and X₄ are N and X₂ and X₃ are C independently        substituted with Y;    -   (iii) X₁ and X₃ are N and X₂ and X₄ are C independently        substituted with Y;    -   (iv) X₂ and X₄ are N and X₁ and X₃ are C independently        substituted with Y;    -   (v) X₁ is N and X₂, X₃, and X₄ are C independently substituted        with Y;    -   (vi) X₃ is N and X₁, X₂ , and X₄ are C independently substituted        with Y;    -   (vii) X₄ is N and X₁, X₂, and X₃ are C independently substituted        with Y;    -   (viii) X₂ is N and X₁, X₃, and X₄ are C independently        substituted with Y; and    -   (ix) X₁, X₂ and X₃ are N and X₄ is C substituted with Y;    -   R1 is H, C₁₋₆ alkylNR5R6, C₁₋₆alkylNR5COR6, C₁₋₆alkylNR5SO₂R6,        C₁₋₆alkylCO₂R5, C₁₋₆alkylCONR5R6, where R5 and R6 are each        independently H, C₁₋₄alkyl, aryl, hetaryl, C₁₋₄alkylaryl,        C₁₋₄alkylhetaryl or may be joined to form an optionally        substituted 3-8 membered ring optionally containing an atom        selected from O, S, NR7;        -   R7 is selected from H, C₁₋₄alkyl;    -   R2 is selected from C₁₋₆alkylOH, OC₂₋₆alkylOH, C₁₋₆alkylNR8R9,        OC₂₋₆alkylNR8R9, C₁₋₆alkylNR8COR9, OC₂₋₆alkylNR8COR9,        C₁₋₆alkylhetaryl, OC₂₋₆alkylhetaryl, OCONR8R9, NR8COOR9,        NR10CONR8R9, CONR8R9, NR8COR12;        -   R8, R9 are each independently H, C₁₋₄alkylNR11R13, hetaryl,            cyclohetalkyl, or may be joined to form an optionally            substituted 3-8 membered ring optionally containing an atom            selected from O, S, NR14;        -   R12 is C₂₋₄alkyl, C₁₋₄alkylNR11R13, hetaryl, cyclohetalkyl;            -   R11, R13 are each independently H, C₁₋₄alkyl, or may be                joined to form an optionally substituted 3-8 membered                ring optionally containing an atom selected from O, S,                NR14;                -   R14 is selected from H, C₁₋₄alkyl;        -   R10 is H, C₁₋₄ alkyl;    -   R3 and R4 are each independently H, halogen, C₁₋₄alkyl, OH,        OC₁₋₄alkyl, CF₃, OCF₃;    -   Q is a bond, or C₁₋₄ alkyl;    -   W is selected from H, C₁₋₄alkyl, C₂₋₆alkenyl; where C₁₋₄alkyl or        C₂₋₆alkenyl may be optionally substituted with C₁₋₄alkyl, OH,        OC₁₋₄alkyl, NR15R16;        -   R15, and R16 are each independently H, C₁₋₄alkyl, C₁₋₄ alkyl            cycloalkyl, C₁₋₄alkyl cyclohetalkyl, aryl, hetaryl, or may            be joined to form an optionally substituted 3-8 membered            ring optionally containing an atom selected from O, S, NR17;            -   R17 is selected from H, C₁₋₄alkyl;    -   A is aryl, hetaryl optionally substituted with 0-3 substituents        independently chosen from halogen, C₁₋₄ alkyl, CF₃, aryl,        hetaryl, OCF₃, OC₁₋₄alkyl, OC₂₋₅alkylNR18R19, Oaryl, Ohetaryl,        CO₂R18, CONR18R19, NR18R19, C₁₋₄ alkylNR18R19,        NR2OC₁₋₄alkylNR18R19, NR18COR19, NR20CONR18R19, NR18SO₂R19;        -   R18, R19 are each independently H, C₁₋₄ alkyl, C₁₋₄ alkyl            cyclohetalkyl, aryl, hetaryl, C₁₋₄alkyl aryl, C₁₋₄ alkyl            hetaryl, or may be joined to form an optionally substituted            3-8 membered ring optionally containing an atom selected            from O, S, NR21;            -   R21 is selected from H, C₁₋₄alkyl;        -   R20 is selected from H, C₁₋₄alkyl;    -   Y is selected from H, C₁₋₄alkyl, OH, NR22R23;        -   R22, R23 are each independently H, C₁₋₄alkyl.

In the above description it will be appreciated that:

-   -   C₁₋₄ alkyl means an unsubstituted or optionally substituted        straight or branched alkyl chain    -   Aryl means unsubstituted or optionally substituted phenyl or        naphthyl.    -   Hetaryl means an unsubstituted or optionally substituted 5- or        6-membered heteroaromatic ring containing one or more        heteroatoms selected from O, N, S.    -   Cycloalkyl means a 3-8 membered saturated ring    -   Cyclohetalkyl means an optionally substituted 3-8 membered        saturated ring containing 1-3 heteroatoms selected from O, S,        NR24, where R24 is H, C₁₋₄ alkyl, aryl, hetaryl.

In a preferred embodiment the compound is selected from compounds of thegeneral formula

or pharmaceutically acceptable prodrugs, salts, hydrates, solvates,crystal forms or diastereomers thereof, wherein:

-   -   X₁, X₂, X₃, X₄ are selected from the following:    -   (i) X₁ and X₂ are N and X₃ and X₄ are C independently        substituted with Y;    -   (ii) X₁ and X₄ are N and X₂ and X₃ are C independently        substituted with Y;    -   (iii) X₁ and X₃ are N and X₂ and X₄ are C independently        substituted with Y;    -   (iv) X₂ and X₄ are N and X₁ and X₃ are C independently        substituted with Y;    -   (v) X₁ is N and X₂, X₃, and X₄ are C independently substituted        with Y;    -   (vi) X₃ is N and X₁, X₂, and X₄ are C independently substituted        with Y;    -   (vii) X₄ is N and X₁, X₂ , and X₃ are C independently        substituted with Y;    -   (viii) X₂ is N and X₁, X₃, and X₄ are C independently        substituted with Y; and    -   (ix) X₁, X₂ and X₃ are N and X₄ is C substituted with Y;    -   R1 is H, C₁₋₆alkyl, C₁₋₆alkylNR5R6, where R5 and R6-are each        independently H, C₁₋₄alkyl, aryl, hetaryl, or may be joined to        form an optionally substituted 3-8 membered ring optionally        containing an atom selected from O, S, NR7;        -   R7 is selected from H, C₁₋₄ alkyl;    -   R2 is selected from C₁₋₆alkylOH, OC₂₋₆alkylOH, C₁₋₆alkylNR8R9,        OC₂₋₆alkylNR8R9, C₁₋₆alkylNR8COR9, OC₂₋₆alkylNR8COR9,        C₁₋₆alkylhetaryl, OC₂₋₆alkylhetaryl, OCONR8R9, NR8COOR9,        NR10CONR8R9, CONR8R9, NR8COR12;        -   R8, R9 are each independently H, C₁₋₄alkyl,            C₁₋₄alkylNR11R13, hetaryl, cyclohetalkyl, or may be joined            to form an optionally substituted 3-8 membered ring            optionally containing an atom selected from O, S, NR14;        -   R12 is C₂₋₄alkyl, C₁₋₄alkylNR11R13, hetaryl, cyclohetalkyl;            -   R11, R13 are each independently H, C₁₋₄alkyl, or may be                joined to form an optionally substituted 3-8 membered                ring optionally containing an atom selected from O, S,                NR14;                -   R14 is selected from H, C₁₋₄alkyl;        -   R10 is H, C₁₋₄alkyl;    -   R3 and R4 are each independently H, halogen, C₁₋₄alkyl, OH,        OC₁₋₄alkyl, CF₃, OCF₃;    -   Q is CH;    -   W is selected from C₁₋₄alkyl, C₂₋₆alkenyl; where C₁₋₄alkyl or        C₂₋₆alkenyl may be optionally substituted with C₁₋₄alkyl, OH,        OC₁₋₄alkyl, NR15R16;        -   R15, and R16 are each independently H, C₁₋₄alkyl, or may be            joined to form an optionally substituted 3-8 membered ring            optionally containing an atom selected from O, S, NR17;            -   R17 is selected from H, C₁₋₄alkyl;    -   A is aryl, hetaryl optionally substituted with 0-2 substituents        independently chosen from halogen, C₁₋₄ alkyl, CF₃, aryl,        hetaryl, OCF₃, OC₁₋₄alkyl; OC₂₋₅alkylNR18R19, Oaryl, Ohetaryl,        CO₂R18, CONR18R19, NR18R19, C₁₋₄ alkylNR18R19,        NR20C₁₋₄alkylNR18R19, NR18COR19, NR20CONR18R19, NR18SO₂R19;        -   R18, R19 are each independently H, C₁₋₄alkyl, C₁₋₄alkyl            cyclohetalkyl, aryl, hetaryl, C₁₋₄alkyl aryl, C₁₋₄alkyl            hetaryl, or may be joined to form an optionally substituted            3-8 membered ring optionally containing an atom selected            from O, S, NR21;            -   R21 is selected from H, C₁₋₄alkyl;        -   R20 is selected from H, C₁₋₄alkyl;    -   Y is selected from H, C₁₋₄alkyl, NR22R23;        -   R22, R23 are each independently H, C₁₋₄alkyl.

In the above description it will be appreciated that:

-   -   C₁₋₄ alkyl means an unsubstituted or optionally substituted        straight or branched alkyl chain    -   Aryl means unsubstituted or optionally substituted phenyl or        naphthyl.    -   Hetaryl means an unsubstituted or optionally substituted 5- or        6-membered heteroaromatic ring containing one or more        heteroatoms selected from O, N, S.    -   Cycloalkyl means a 3-8 membered saturated ring    -   Cyclohetalkyl means an optionally substituted 3-8 membered        saturated ring containing 1-3 heteroatoms selected from O, S,        NR24, where R24 is H, C₁₋₄ alkyl, aryl, hetaryl.

In a sixth aspect, the present invention provides a compositioncomprising a carrier and one or more compounds of the fifth aspect.

In a seventh aspect, the present invention provides a method oftreatment of a hyperproliferation-related disorder in a subject, themethod comprising administering a therapeutically effective amount of atleast one of the compounds of the fifth aspect or a compositionaccording to the sixth aspect.

Preferably, the hyperproliferation-related disorder is treatable by themodulation of microtubule polymerisation.

In an eighth aspect, the present invention consists in a method oftreating a protein-kinase related disorder in a subject, the methodcomprising administering a therapeutically effective amount of at leastone of the compounds of the fifth aspect or a composition according tothe sixth aspect.

In a ninth aspect, the present invention provides a compound of thegeneral formula

or pharmaceutically acceptable prodrugs, salts, hydrates, solvates,crystal forms or diastereomers thereof, wherein:

-   -   X₁, X₂, X₃, X₄ are selected from the following:    -   (i) X₁ and X₂ are N and X₃ and X₄ are C independently        substituted with Y;    -   (ii) X₁ and X₄ are N and X₂ and X₃ are C independently        substituted with Y;    -   (iii) X₂ and X₄ are N and X₁ and X₃ are C independently        substituted with Y;    -   (iv) X₁ is N and X₂, X₃, and X₄ are C independently substituted        with Y;    -   (v) X₃ is N and X₁, X₂ and X₄ are C independently substituted        with Y;    -   (vi) X₄ is N and X₁, X₂, and X₃ are C independently substituted        with Y;    -   (vii) X₂ is N and X₁, X₃, and X₄ are C independently substituted        with Y; and    -   (viii) X₁, X₂ and X₃ are N and X₄ is C substituted with Y;    -   R1 is H, C₁₋₆alkyl, C₁₋₆ alkylNR5R6, C₁₋₆alkylNR5COR6,        C₁₋₆alkylNR5SO₂R6, C₁₋₆alkylCO₂R5, C₁₋₆alkylCONR5R6, where R5        and R6 are each independently H, C₁₋₄alkyl, aryl, hetaryl,        C₁₋₄alkylaryl, C₁₋₄alkylhetaryl or may be joined to form an        optionally substituted 3-8 membered ring optionally containing        an atom selected from O, S, NR7;        -   R7 is selected from H, C₁₋₄ alkyl;    -   R2 is selected from OH, OC₁₋₆alkyl, C₁₋₆alkylOH, OC₂₋₆alkylOH,        C₁₋₆alkylNR8R9, OC₂₋₆alkylNR8R9, C₁₋₆alkylNR8COR9,        OC₂₋₆alkylNR8COR9, C₁₋₆alkylhetaryl, OC₂₋₆alkylhetaryl,        OCONR8R9, NR8COOR9, NR10CONR8R9, CONR8R9, NR8COR12;        -   R8, R9 are each independently H, C₁₋₄alkyl,            C₁₋₄alkylNR11R13, hetaryl, cyclohetalkyl, or may be joined            to form an optionally substituted 3-8 membered ring            optionally containing an atom selected from O, S, NR14;        -   R12 is C₂₋₄alkyl, C₁₋₄alkylNR11R13, hetaryl, cyclohetalkyl;            -   R11, R13 are each independently H, C₁₋₄alkyl, or may be                joined to form an optionally substituted 3-8 membered                ring optionally containing an atom selected from O, S,                NR14;                -   R14 is selected from H, C₁₋₄alkyl;        -   R10 is H, C₁₋₄ alkyl;    -   R3 and R4 are each independently H, halogen, C₁₋₄alkyl, OH,        OC₁₋₄alkyl, CF₃, OCF₃;    -   Q is a bond, or C₁₋₄alkyl;    -   W is selected from H, C₁₋₄alkyl, C₂₋₆alkenyl; where C₁₋₄alkyl or        C₂₋₆alkenyl may be optionally substituted with C₁₋₄alkyl, OH,        OC₁₋₄alkyl, NR15R16;        -   R15, and R16 are each independently H, C₁₋₄alkyl, C₁₋₄alkyl            cycloalkyl, C₁₋₄alkyl cyclohetalkyl, aryl, hetaryl, or may            be joined to form an optionally substituted 3-8 membered            ring optionally containing an atom selected from O, S, NR17;            -   R17 is selected from H, C₁₋₄alkyl;    -   A is aryl, hetaryl optionally substituted with 0-3 substituents        independently chosen from halogen, C₁₋₄ alkyl, CF₃, aryl,        hetaryl, OCF₃, OC₁₋₄alkyl, OC₂₋₅alkylNR18R19, Oaryl, Ohetaryl,        CO₂R18, CONR18R19, NR18R19, C₁₋₄ alkylNR18R19,        NR20C₁₋₄alkylNR18R19, NR18COR19, NR20CONR18R19, NR18SO₂R19;        -   R18, R19 are each independently H, C₁₋₄ alkyl, C₁₋₄ alkyl            cyclohetalkyl, aryl, hetaryl, C₁₋₄alkyl aryl, C₁₋₄ alkyl            hetaryl, or may be joined to form an optionally substituted            3-8 membered ring optionally containing an atom selected            from O, S, NR21;            -   R21 is selected from H, C₁₋₄ alkyl;        -   R20 is selected from H, C₁₋₄ alkyl;    -   Y is selected from H, C₁₋₄alkyl, OH, NR22R23;        -   R22, R23 are each independently H, C₁₋₄ alkyl.

In the above description it will be appreciated that:

-   -   C₁₋₄ alkyl means an unsubstituted or optionally substituted        straight or branched alkyl chain    -   Aryl means unsubstituted or optionally substituted phenyl or        naphthyl.    -   Hetaryl means an unsubstituted or optionally substituted 5- or        6-membered heteroaromatic ring containing one or more        heteroatoms selected from O, N, S.    -   Cycloalkyl means a 3-8 membered saturated ring    -   Cyclohetalkyl means an optionally substituted 3-8 membered        saturated ring containing 1-3 heteroatoms selected from O, S,        NR24, where R24 is H, C₁₋₄ alkyl, aryl, hetaryl.

In a tenth aspect, the present invention provides a compositioncomprising a carrier and one or more compounds of the ninth aspect.

In an eleventh aspect, the present invention provides a method oftreatment of a hyperproliferation-related disorder in a subject, themethod comprising administering a therapeutically effective amount of atleast one of the compounds of the ninth aspect or a compositionaccording to the tenth aspect.

Preferably, the hyperproliferation-related disorder is treatable by themodulation of microtubule polymerisation.

In a twelfth aspect, the present invention consists in a method oftreating a protein-kinase related disorder in a subject, the methodcomprising administering a therapeutically effective amount of at leastone of the compounds of the ninth aspect or a composition according tothe tenth aspect.

The compounds of this invention include all conformational isomers (eg.cis and trans isomers). The compounds of the present invention haveasymmetric centers and therefore exist in different enantiomeric anddiastereomeric forms. This invention relates to the use of all opticalisomers and stereoisomers of the compounds of the present invention, andmixtures thereof, and to all pharmaceutical compositions and methods oftreatment that may employ or contain them. The compounds of formula I,II, III, IV and V may also exist as tautomers. This invention relates tothe use of all such tautomers and mixtures thereof.

Compounds of formula I, II, III, IV and V having free amino, amido,hydroxy or carboxylic groups can be converted into prodrugs. Prodrugsinclude compounds wherein an amino acid residue, or a polypeptide chainof two or more (eg, two, three or four) amino acid residues which arecovalently joined through peptide bonds to free amino, hydroxy andcarboxylic acid groups of compounds of formulae I, II, III, IV and V.The amino acid residues include the 20 naturally occurring amino acidscommonly designated by three letter symbols and also include,4-hydroxyproline, hydroxylysine, demosine, isodemosine,3-methylhistidine, norvaline, beta-alanine, gamma-aminobutyric acid,citrulline, homocysteine, homoserine, ornithine and methioine sulfone.Prodrugs also include compounds wherein carbonates, carbamates, amidesand alkyl esters which are covalently bonded to the above substituentsof formulae I, II, III, IV and V through the carbonyl carbon prodrugsidechain. Prodrugs also include phosphate derivatives of compounds offormulae I, II, III, IV and V (such as acids, salts of acids, or esters)joined through a phosphorus-oxygen bond to a free hydroxyl of compoundsof formulae I, II, III, IV and V. Prodrugs also include compoundswherein acyloxyalkyl or phosphonooxyalkyl moieties are covalentlyattached to compounds of formula I, II, III, IV and V possessing a freehydroxyl group. Acyloxyalkyl or phosphonooxyalkyl moieties may also becovalently attached to compounds of formulae I, II, III, IV and Vpossessing a pyridyl ring through formation of a N-(acyloxyalkyl)- orN-(phosphonooxyalkyl)-pyridinium salt. This invention also encompassespharmaceutical compositions containing prodrugs of compounds of theformula I, II, III, IV and V. In addition, this invention encompassesmethods of treating or preventing disorders that can be treated orprevented by the modulation of microtubule dynamics comprisingadministering prodrugs of compounds of the formula I, II, III, IV and V.This invention further encompasses methods of treating or preventingdisorders that can be treated or prevented by the disruption ofmicrotubules comprising administering prodrugs of compounds of theformula II, III, IV and V.

In a still further preferred embodiment the compound possesses Schirality at the chiral carbon bearing W, where W is C₁₋₄ alkyl orC₁₋₄alkylamino. The compound can be used as a purified isomer or as amixture of any ratio of isomers. It is however preferred that themixture comprises at least 70%, 80%, 90%, 95%, or 99% of the preferredisomer.

In a yet further preferred embodiment, the present invention provides acompound selected from the compounds of Table 1; compositions comprisinga carrier and one or more compounds of Table 1; and, a method oftreatment of a hyperproliferation-related disorder in a subject, themethod comprising administering at least one of the compounds of Table1.

More preferably, the hyperproliferation related disorder is treatable bythe modulation of microtubule polymerisation.

The present invention further provides for a method of modulatingmicrotubule polymerisation in a subject, the method comprising theadministration of a therapeutically effective amount of a compound ofany one of Formulas I, II, III, IV and V.

In a preferred embodiment, there is provided a method of modulatingmicrotubule polymerisation in a subject, the method comprising theadministration of a therapeutically effective amount of a compoundselected from the compounds of Table 1.

The present invention provides that the compounds of formula I, II, III,IV and V may be administered for the treatment ofhyperproliferation-related disorders. Preferably, thehyperproliferation-related disorder is treatable by the modulation ofmicrotubule polymerisation. In a preferred embodiment, thehyperproliferation-related disorder or disease state is selected fromthe group consisting of Cancer, such as fibrosarcoma, myxosarcoma,liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma,endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma,synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma,rhabdomyosarcoma, colon carcinoma, pancreatic cancer, breast cancer,ovarian cancer, prostate cancer, squamous cell carcinoma, basal cellcarcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous glandcarcinoma, papillary carcinoma, papillary adenocarcinomas,cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renalcell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma,seminoma, embryonal carcinoma, Wilms' tumor, cervical cancer, testiculartumor, lung carcinoma, small cell lung carcinoma, bladder carcinoma,epithelial carcinoma, glioma, astrocytoma, medulloblastoma,craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acousticneuroma, oligodendroglioma, meningioma, melanoma, neuroblastoma, andretinoblastoma, and carcinomas forming from tissue of the breast,prostate, kidney, bladder or colon, and neoplastic disorders arising inadipose tissue, such as adipose cell tumors, e.g., lipomas,fibrolipomas, lipoblastomas, lipomatosis, hibemomas, hemangiomas and/orliposarcomas; infectious diseases such as viral, malarial and bacterialinfections; vascular restenosis; inflammatory diseases, such asautoimmune diseases, glomerular nephritis myocardial infarction andpsoriasis.

The compounds of formula II, II and IV may also be used in the treatmentof a hyperproliferation-related disorder or disease state and/or aprotein-kinase related disorder or disease state. Preferably, thedisease state is selected from the group consisting of Atopy, such asAllergic Asthma, Atopic Dermatitis (Eczema), and Allergic Rhinitis; CellMediated Hypersensitivity, such as Allergic Contact Dermatitis andHypersensitivity Pneumonitis; Rheumatic Diseases, such as Systemic LupusErythematosus (SLE), Rheumatoid Arthritis, Juvenile Arthritis, Sjögren'sSyndrome, Scleroderma, Polymyositis, Ankylosing Spondylitis, PsoriaticArthritis; Other autoimmune diseases such as Type I diabetes, autoimmunethyroid disorders, and Alzheimer's disease; Viral Diseases, such asEpstein Barr Virus (EBV), Hepatitis B, Hepatitis C, HIV, HTLV 1,Varicella-Zoster Virus (VZV), Human Papilloma Virus (HPV).

In the treatment or prevention of hyperproliferation-related disordersand/or protein kinase related disorders an appropriate dosage level willgenerally be about 0.01 to 500 mg per kg patient body weight per daywhich can be administered in single or multiple doses. Preferably, thedosage level will be about 0.1 to about 250 mg/kg per day; morepreferably about 0.5 to about 100 mg/kg per day. A suitable dosage levelmay be about 0.01 to 250 mg/kg per day, about 0.05 to 100 mg/kg per day,or about 0.1 to 50 mg/kg per day. Within this range the dosage may be0.05 to 0.5, 0.5 to 5 or 5 to 50 mg/kg per day. For oral administration,the compositions are preferably provided in the form of tabletscontaining 1.0 to 1000 milligrams of the active ingredient, particularly1.0, 5.0, 10.0, 15.0. 20.0, 25.0, 50.0, 75.0, 100.0, 150.0, 200.0,250.0, 300.0, 400.0, 500.0, 600.0, 750.0, 800.0, 900.0, and 1000.0milligrams of the active ingredient for the symptomatic adjustment ofthe dosage to the patient to be treated. The compounds may beadministered on a regimen of 1 to 4 times per day, preferably once ortwice per day.

It will be understood, however, that the specific dose level andfrequency of dosage for any particular patient may be varied and willdepend upon a variety of factors including the activity of the specificcompound employed, the metabolic stability and length of action of thatcompound, the age, body weight, general health, sex, diet, mode and timeof administration, rate of excretion, drug combination, the severity ofthe particular condition, and the host undergoing therapy.

In addition to primates, such as humans, a variety of other mammals canbe treated according to the method of the present invention. Forinstance, mammals including, but not limited to, cows, sheep, goats,horses, dogs, cats, guinea pigs, rats or other bovine, ovine, equine,canine, feline, rodent or murine species can be treated. However, themethod can also be practiced in other species, such as avian species(e.g., chickens).

Diseases and conditions associated with inflammation and infection canbe treated using the method of the present invention. In a preferredembodiment, the disease or condition is one in which the actions ofeosinophils and/or lymphocytes are to be inhibited or promoted, in orderto modulate the inflammatory response.

The subjects treated in the above methods, in whom which cell growthinhibition is desired, are mammals, including, but not limited to, cows,sheep, goats, horses, dogs, cats, guinea pigs, rats or other bovine,ovine, equine, canine, feline, rodent or murine species, and preferablya human being, male or female.

The term “therapeutically effective amount” means the amount of thesubject composition that will elicit the biological or medical responseof a tissue, system, animal or human that is being sought by theresearcher, veterinarian, medical doctor or other clinician.

The terms “administration of” and or “administering a” compound shouldbe understood to mean providing a compound of the invention to theindividual in need of treatment.

The present invention provides pharmaceutical compositions comprising atleast one of the compounds of the formula I, II, III, IV and V capableof treating a hyperproliferation-related disorder in an amount effectivetherefore, and a pharmaceutically acceptable vehicle or diluent. Thecompositions of the present invention may contain other therapeuticagents as described below, and may be formulated, for example, byemploying conventional solid or liquid vehicles or diluents, as well aspharmaceutical additives of a type appropriate to the mode of desiredadministration (for example, excipients, binders, preservatives,stabilizers, flavors, etc.) according to techniques such as those wellknown in the art of pharmaceutical formulation.

The compounds of the formula I, II, III, IV and V may be administered byany suitable means, for example, orally, such as in the form of tablets,capsules, granules or powders; sublingually; buccally; parenterally,such as by subcutaneous, intravenous, intramuscular, or intracisternalinjection or infusion techniques (e.g., as sterile injectable aqueous ornon-aqueous solutions or suspensions); nasally such as by inhalationspray; topically, such as in the form of a cream or ointment; orrectally such as in the form of suppositories; in dosage unitformulations containing non-toxic, pharmaceutically acceptable vehiclesor diluents. The compounds may, for example, be administered in a formsuitable for immediate release or extended release. Immediate release orextended release may be achieved by the use of suitable pharmaceuticalcompositions comprising the present compounds, or, particularly in thecase of extended release, by the use of devices such as subcutaneousimplants or osmotic pumps.

The term “composition” as used herein is intended to encompass a productcomprising the specified ingredients in the specified amounts, as wellas any product which results, directly or indirectly, from combinationof the specified ingredients in the specified amounts. By“pharmaceutically acceptable” it is meant the carrier, diluent orexcipient must be compatible with the other ingredients of theformulation and not deleterious to the recipient thereof.

The pharmaceutical compositions for the administration of the compoundsof this invention may conveniently be presented in dosage unit form andmay be prepared by any of the methods well known in the art of pharmacy.All methods include the step of bringing the active ingredient intoassociation with the carrier which constitutes one or more accessoryingredients. In general, the pharmaceutical compositions are prepared byuniformly and intimately bringing the active ingredient into associationwith a liquid carrier or a finely divided solid carrier or both, andthen, if necessary, shaping the product into the desired formulation. Inthe pharmaceutical composition the active object compound is included inan amount sufficient to produce the desired effect upon the process orcondition of diseases. As used herein, the term “composition” isintended to encompass a product comprising the specified ingredients inthe specified amounts, as well as any product which results, directly orindirectly, from combination of the specified ingredients in thespecified amounts.

The pharmaceutical compositions containing the active ingredient may bein a form suitable for oral use, for example, as tablets, troches,lozenges, aqueous or oily suspensions, dispersible powders or granules,emulsions, hard or soft capsules, or syrups or elixirs. Compositionsintended for oral use may be prepared according to any method known tothe art for the manufacture of pharmaceutical compositions and suchcompositions may contain one or more agents selected from the groupconsisting of sweetening agents, flavoring agents, coloring agents andpreserving agents in order to provide pharmaceutically elegant andpalatable preparations. Tablets contain the active ingredient inadmixture with non-toxic pharmaceutically acceptable excipients whichare suitable for the manufacture of tablets. These excipients may be forexample, inert diluents, such as calcium carbonate, sodium carbonate,lactose, calcium phosphate or sodium phosphate; granulating anddisintegrating agents, for example, corn starch, or alginic acid;binding agents, for example starch, gelatin or acacia, and lubricatingagents, for example magnesium stearate, stearic acid or talc. Thetablets may be uncoated or they may be coated by known techniques todelay disintegration and absorption in the gastrointestinal tract andthereby provide a sustained action over a longer period. For example, atime delay material such as glyceryl monostearate or glyceryl distearatemay be employed. They may also be coated to form osmotic therapeutictablets for control release.

Formulations for oral use may also be presented as hard gelatin capsuleswherein the active ingredient is mixed with an inert solid diluent, forexample, calcium carbonate, calcium phosphate or kaolin, or as softgelatin capsules wherein the active ingredient is mixed with water or anoil medium, for example peanut oil, liquid paraffin, or olive oil.

Aqueous suspensions contain the active materials in admixture withexcipients suitable for the manufacture of aqueous suspensions. Suchexcipients are suspending agents, for example sodiumcarboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose,sodium alginate, polyvinyl-pyrrolidone, gum tragacanth and gum acacia;dispersing or wetting agents may be a naturally-occurring phosphatide,for example lecithin, or condensation products of an alkylene oxide withfatty acids, for example polyoxyethylene stearate, or condensationproducts of ethylene oxide with long chain aliphatic alcohols, forexample heptadecaethyleneoxycetanol, or condensation products ofethylene oxide with partial esters derived from fatty acids and ahexitol such as polyoxyethylene sorbitol monooleate, or condensationproducts of ethylene oxide with partial esters derived from fatty acidsand hexitol anhydrides, for example polyethylene sorbitan monooleate.The aqueous suspensions may also contain one or more preservatives, forexample ethyl, or n-propyl, p-hydroxybenzoate, one or more coloringagents, one or more flavoring agents, and one or more sweetening agents,such as sucrose or saccharin.

Oily suspensions may be formulated by suspending the active ingredientin a vegetable oil, for example arachis oil, olive oil, sesame oil orcoconut oil, or in a mineral oil such as liquid paraffin. The oilysuspensions may contain a thickening agent, for example beeswax, hardparaffin or cetyl alcohol. Sweetening agents such as those set forthabove, and flavoring agents may be added to provide a palatable oralpreparation. These compositions may be preserved by the addition of ananti-oxidant such as ascorbic acid.

Dispersible powders and granules suitable for preparation of an aqueoussuspension by the addition of water provide the active ingredient inadmixture with a dispersing or wetting agent, suspending agent and oneor more preservatives. Suitable dispersing or wetting agents andsuspending agents are exemplified by those already mentioned above.Additional excipients, for example sweetening, flavoring and coloringagents, may also be present.

The pharmaceutical compositions of the invention may also be in the formof oil-in-water emulsions. The oily phase may be a vegetable oil, forexample olive oil or arachis oil, or a mineral oil, for example liquidparaffin or mixtures of these. Suitable emulsifying agents may benaturally-occurring gums, for example gum acacia or gum tragacanth,naturally-occurring phosphatides, for example soy bean, lecithin, andesters or partial esters derived from fatty acids and hexitolanhydrides, for example sorbitan monooleate, and condensation productsof the said partial esters with ethylene oxide, for examplepolyoxyethylene sorbitan monooleate. The emulsions may also containsweetening and flavoring agents.

Syrups and elixirs may be formulated with sweetening agents, for exampleglycerol, propylene glycol, sorbitol or sucrose. Such formulations mayalso contain a demulcent, a preservative and flavoring and coloringagents.

The pharmaceutical compositions may be in the form of a sterileinjectable aqueous or oleagenous suspension. This suspension may beformulated according to the known art using those suitable dispersing orwetting agents and suspending agents which have been mentioned above.The sterile injectable preparation may also be a sterile injectablesolution or suspension in a non-toxic parenterally-acceptable diluent orsolvent, for example as a solution in 1,3-butane diol. Among theacceptable vehicles and solvents that may be employed are water,Ringer's solution and isotonic sodium chloride solution. In addition,sterile, fixed oils are conventionally employed as a solvent orsuspending medium. For this purpose any bland fixed oil may be employedincluding synthetic mono- or diglycerides. In addition, fatty acids suchas oleic acid find use in the preparation of injectables.

The compounds of the present invention may also be administered in theform of suppositories for rectal administration of the drug. Thesecompositions can be prepared by mixing the drug with a suitablenon-irritating excipient which is solid at ordinary temperatures butliquid at the rectal temperature and will therefore melt in the rectumto release the drug. Such materials are cocoa butter and polyethyleneglycols.

For topical use, creams, ointments, jellies, solutions or suspensions,etc., containing the compounds of the present invention are employed.(For purposes of this application, topical application shall includemouthwashes and gargles.)

The compounds of the present invention can also be administered in theform of liposomes. As is known in the art, liposomes are generallyderived from phospholipids or other lipid substances. Liposomes areformed by mono- or multilamellar hydrated liquid crystals that aredispersed in an aqueous medium. Any non-toxic, physiologicallyacceptable and metabolisable lipid capable of forming liposomes can beused. The present compositions in liposome form can contain, in additionto a compound of the present invention, stabilisers, preservatives,excipients and the like. The preferred lipids are the phospholipids andphosphatidyl cholines, both natural and synthetic. Methods to formliposomes are known in the art.

The pharmaceutical composition and method of the present invention mayfurther comprise other therapeutically active compounds as noted hereinwhich are usually applied in the treatment of the above mentionedpathological conditions. Selection of the appropriate agents for use incombination therapy may be made by one of ordinary skill in the art,according to conventional pharmaceutical principles. The combination oftherapeutic agents may act synergistically to effect the treatment orprevention of the various disorders described above. Using thisapproach, one may be able to achieve therapeutic efficacy with lowerdosages of each agent, thus reducing the potential for adverse sideeffects.

Examples of other therapeutic agents include the following:

Tyrosine kinase inhibitors, such as imatinib (Glivec™), and gefitinib(Iressa™) inter alia; cyclosporins (e.g., cyclosporin A), CTLA4-Ig,antibodies such as ICAM-3, anti-IL-2 receptor (Anti-Tac), anti-CD45RB,anti-CD2, anti-CD3 (OKT-3), anti-CD4, anti-CD80, anti-CD86, agentsblocking the interaction between CD40 and gp39, such as antibodiesspecific for CD40 and/or gp39 (i.e., CD154), fusion proteins constructedfrom CD40 and gp39 (CD401g and CD8gp39), inhibitors, such as nucleartranslocation inhibitors, of NF-kappa B function, such asdeoxyspergualin (DSG), cholesterol biosynthesis inhibitors such as HMGCoA reductase inhibitors (lovastatin and simvastatin), non-steroidalantiinflammatory drugs (NSAIDs) such as ibuprofen, aspirin,acetaminophen and cyclooxygenase inhibitors such as rofecoxib, steroidssuch as prednisolone or dexamethasone, gold compounds, antiproliferativeagents such as methotrexate, FK506 (tacrolimus, Prograf), mycophenolatemofetil, antineoplastic agents such as azathioprine, VP-16, etoposide,fludarabine, cisplatin, doxorubicin, adriamycin, amsacrine,camptothecin, cytarabine, gemcitabine, fluorodeoxyuridine, melphalan andcyclophosphamide, TNF-α inhibitors such as tenidap, anti-TNF antibodiesor soluble TNF receptor, and rapamycin (sirolimus or Rapamune) orderivatives thereof.

When other therapeutic agents are employed in combination with thecompounds of the present invention they may be used for example inamounts as noted in the Physician Desk Reference (PDR) or as otherwisedetermined by one of ordinary skill in the art.

The pharmaceutical composition and method of the present invention mayfurther comprise other therapeutically active compounds as noted hereinwhich are known inhibitors or substrates of drug efflux systems or drugdetoxification and excretory systems. Such systems includeP-glycoprotein, multidrug resistance-associated protein, lung resistanceprotein and glutathione S-transferase isoenzymes alpha, mu, pi, sigma,theta, zeta and kappa. Co-administration of drugs known to inhibit orreduce the activity of these systems may increase the efficacy of thecompounds described in the present invention through increasing theamount of therapeutic agent in the cell. Using this approach, one may beable to achieve therapeutic efficacy with lower dosages, thus reducingthe potential for adverse side effects. Examples of inhibitors orsubstrates for these systems include; verapamil, probenecid,dipyridamole, ethacrynic acid, indomethacin, sulfasalazine, buthioninesulfoximine, cyclosporin A and tamoxifen.

Throughout this specification the word “comprise”, or variations such as“comprises” or “comprising”, will be understood to imply the inclusionof a stated element, integer or step, or group of elements, integers orsteps, but not the exclusion of any other element, integer or step, orgroup of elements, integers or steps.

All publications mentioned in this specification are herein incorporatedby reference.

Any discussion of documents, acts, materials, devices, articles or thelike which has been included in the present specification is solely forthe purpose of providing a context for the present invention. It is notto be taken as an admission that any or all of these matters form partof the prior art base or were common general knowledge in the fieldrelevant to the present invention as it existed in Australia before thepriority date of each claim of this application.

In order that the nature of the present invention may be more clearlyunderstood preferred forms thereof will now be described by reference tothe following non-limiting Examples.

EXAMPLES

Materials and Methods:

Compound Synthesis

Compounds are generally prepared in a 2-step process starting from adihaloheterocycle.

The first step is a nucleophilic aromatic substitution to generate amonoamino-monohalo intermediate.

The nucleophilic aromatic substitution is typically carried out byaddition of a primary or secondary amine to the di-halogenatedheterocycle in a solvent such as ethanol, isopropanol, tert-butanol,dioxane, THF, DMF, ethoxyethanol, toluene or xylene. The reaction istypically performed at elevated temperature in the presence of excessamine or a non-nucleophilic base such as triethylamine ordiisopropylethylamine, or an inorganic base such as potassium carbonateor sodium carbonate.

Alternatively, the amino substituent may be introduced through atransition metal catalysed amination reaction. Typical catalysts forsuch transformations include Pd(OAc)₂/P(t-Bu)₃, Pd₂(dba)₃/BINAP andPd(OAc)₂/BINAP. These reactions are typically carried out in solventssuch as toluene or dioxane, in the presence of bases such as caesiumcarbonate or sodium or potassium tert-butoxide at temperatures rangingfrom room temperature to reflux.

The amines employed in the first step of the synthesis of thesecompounds are obtained commercially or are prepared using methods wellknown to those skilled in the art. Of particular interest areα-alkylbenzylamines which may be prepared through reduction of oximes.Typical reductants include lithium aluminium hydride, hydrogen gas inthe presence of palladium on charcoal catalyst, Zn in the presence ofhydrochloric acid, sodium borohydride in the presence of a Lewis acidsuch as TiCl₃, ZrCl₄, NiCl₂ and MoO₃, or sodium borohydride inconjunction with Amberlyst H15 ion exchange resin and LiCl.

α-Alkylbenzylamines may also be prepared by reductive amination of thecorresponding ketones. A classical method for such a transformation isthe Leuckart-Wallach reaction, though catalytic conditions (HCO₂NH₄,[(CH₃)₅C₅RhCl₂]₂) or alternative procedures (e.g. NH₄OAc, Na(CN)BH₃) canalso be used.

α-Alkylbenzylamines may also be prepared from the correspondingα-alkylbenzyl alcohols. Such methods include derivatisation of thehydroxyl as a mesylate or tosylate and displacement with a nitrogennucleophile, such as phthalimide or azide which is converted to theprimary amine using conventional synthetic methods; or, displacement ofthe hydroxyl with a suitable nitrogen nucleophile under Mitsunobu-likeconditions. α-Alkylbenzyl alcohols can be prepared by reduction of thecorresponding ketones with a reducing agent such as sodium borohydridein a solvent such as methanol. Alternatively, α-alkylbenzyl alcohols canbe obtained through addition of an alkyl metal species (such as aGrignard reagent) to a benzaldehyde derivative, typically performed atroom temperature or below in solvents such as tetrahydrofuran.

α-Alkyl benzylamines of high optical purity may be prepared from chiralα-alkyl benzyl alcohols using the methods outlined above. The chiralα-alkyl benzyl alcohols may be obtained through chiral reduction of thecorresponding ketones. Chiral reducing methods are now well known inorganic chemistry and include enzymatic processes, asymmetrichydrogenation procedures and chiral oxazaborolidines.

The second step of the synthesis typically involves a palladium mediatedcross-coupling of the monoamino-monochloro intermediate with a suitablyfunctionalised coupling partner. Typical coupling partners are boronicacids or esters (Suzuki coupling: see for example Miyaura, N. andSuzuki, Chem Rev. 1995, 952457), stannanes (Stille coupling: see forexample Stille, J. K., Angew. Chem., Int. Ed. Engl., 1986, 25, 508),Grignard reagents (Kumada coupling: Kumada, M.; Tamao, K.; Sumitani, K.Org. Synth. 1988, Coll. Vol. 6, 407.) or organozinc species (Negishicoupling: Negishi, E.; J. Organomet. Chem. 2002, 653, 34).

The Suzuki coupling is the preferred coupling method and is typicallyperformed in a solvent such as DME, THF, DMF, ethanol, propanol,toluene, or 1,4-dioxane in the presence of a base such as potassiumcarbonate, lithium hydroxide, caesium carbonate, sodium hydroxide,potassium fluoride or potassium phosphate. The reaction may be carriedout at elevated temperatures and the palladium catalyst employed may beselected from Pd(PPh₃)₄, Pd(OAc)₂, [PdCl₂(dppf)], Pd₂(dba)₃/P(t-Bu)₃.

The products formed from this reaction sequence may be furtherderivatised using techniques well-known to those skilled in the art.Alternatively, derivatisation of the mono-amino mono-halo intermediatemay be undertaken prior to displacement of the halo substituent. Thisderivatisation typically involves functionality originally present onthe amine species and employs methods well known to those skilled in theart.

Alternatively the preparation sequence can be reversed beginning withthe palladium mediated cross-coupling reaction to furnish a mono-haloheterocyclic species. Amine displacement of the halo substituent canthen be undertaken using procedures outlined above.

In order that the nature of the present invention may be more clearlyunderstood, preferred forms thereof will now be described with referenceto the following non-limiting examples.

Example 1 1-[4-(Trifluoromethyl)phenyl]butan-1-ol

A 2M solution of propylmagnesium chloride in ether (4 ml, 8 mmol) wasadded to a solution of the aldehyde (1.14 g, 6.6 mmol) in dry THF (10ml) cooled to 0° C. under N₂. The mixture was stirred for 16 h at roomtemperature, after which time saturated ammonium chloride solution wasadded. The product was extracted into ethyl acetate, and the ethylacetate layer dried and concentrated to furnish pure product (1.4 g,98%).

1H-n.m.r. (CDCl₃) δ 0.94 (t, 3H, J=7.2 Hz, CH₃), 1.41 (m, 2H, CH₂), 1.75(m, CH₂, 2H), 4.77 (br s, 1H, CH), 7.44-7.62 (m, 4H, ArH)

Example 2 1-(1-Azidobutyl)-4-(trifluoromethyl)benzene

A solution of 1-[4-(trifluoromethyl)phenyl]butan-1-ol (1.4 g, 6.4 mmol)and diphenylphosphoryl azide (2.8 ml, 12.8 mmol) in THF (6 mL) cooled to−10° C. under N₂ was treated with DBU (1.9 mL, 12.8 mmol). The resultingsolution was stirred at room temperature for 20 hours and then dilutedwith a mixture of ether and H₂O. The organic phase was dried andconcentrated and the residue purified by column chromatography usinghexane:ethyl acetate (10:1) as eluent to furnish pure azide (0.85 g,54%).

¹H-n.m.r. (CDCl₃) δ 0.94 (t, 3H, J=7.2 Hz, CH₃), 1.37 (m, 2H, CH₂), 1.75(m, 2H, CH₂), 4.50 (t, 1H, CH), 7.42 (d, J=7.8 Hz, 2H, ArH), 7.64 (d,2H, J=7.8 Hz, ArH)

Example 3 1-[4-(Trifluoromethyl)phenyl]butan-1-amine

A mixture of 1-(1-azidobutyl)-4-(trifluoromethyl)benzene (0.84 g, 3.5mmol) and triphenylphosphine (1.8 g, 6.9 mmol) in ethyl acetate (6 mL)and 10% HCl (6 ml) was stirred at room temperature for 64 h. The aqueousphase was collected and the organic phase extracted with 10% HCl (3×5mL). The aqueous layers were combined and basified with 5M NaOH, andthen extracted with ethyl acetate (5×15 mL). The organic phase was driedand concentrated to give pure amine (0.4 g, 54%).

¹H-n.m.r. (CDCl₃) δ 0.91 (t, J=7.4 Hz, 3H CH₃), 1.31 (m, 2H, CH₂), 1.62(m, 2H, CH₂), 3.97 (m, 1H, CH), 7.43 (AA′XX′, 2H, ArH), 7.58 (AA′XX′,2H, ArH)

Example 4 3-Fluoro-N-methoxy-N-methylbenzamide

To a suspension of 3-fluorobenzoic acid (140 mg, 1 mmol) andN,O-dimethylhydroxylamine hydrochloride (107 mg, 1.1 mmol) indichloromethane (2.5 mL) was added1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (EDC) (211mg, 1.1 mmol) and the mixture stirred at room temperature for 75 h. Thesolvents were removed under reduced pressure and the residuechromatographed using ethyl acetate-hexane (4:6) to separate the pureproduct (130 mg, 71%).

¹H-n.m.r. (CDCl₃) δ 3.36 (s, 3H, N-Me), 3.55 (s, 3H, N—OMe), 7.1.-7.2(m, 1H, ArH), 7.3-7.5 (m, 3H, Ar)

Example 5 1-(3-Fluorophenyl)butan-1-one

To a solution of 3-fluoro-N-methoxy-N-methylbenzamide (130 mg, 0.71mmol) in dry THF (2 mL) cooled to −10° C. was added propyl magnesiumchloride (532 μl, 2M solution in ether, 1.1 mmol) under nitrogen. Thesolution was stirred at −10° C. for 1 h and at room temperature for 75min. The solution was the poured into saturated aqueous ammoniumchloride and the product extracted into ethyl acetate (3×25 mL). Theorganic layers were combined, washed with brine, dried (MgSO₄) andconcentrated to give a pale yellow oil, which was purified bychromatography using ethyl acetate-hexane (1:9) to separate the pureproduct (78 mg, 66%).

¹H-n.m.r. (CDCl₃) δ 1.01 (t, 3H, J=7.4 Hz, Me), 1.77 (m, 2H, CH₂ CH₃),2.93 (t, J=7.2 Hz, 2H, COCH₂), 7.15-7.30 (m, 1H, Ar), 7.35-7.50 (m, 1H,Ar), 7.60-7.70 (m, 1H, Ar), 7.70-7.80 (m, 1H, Ar).

Example 6 1-(5-Methylpyridin-3-yl)butan-1-amine

To a solution of 1-(5-methylpyridin-3-yl)butan-1-one (800 mg, 4.9 mmol)and ammonium formate (1.55 g, 24.5 mmol) in methanol (5 mL) undernitrogen was added dichloro(pentamethylcyclopentadienyl)rhodium (LH)dimer (45 mg, 0.074 mmol). The solution was heated at reflux for 8 h.after which time the solution was cooled to room temperature andacidified to pH ˜2 with 2M HCl. The mixture was washed withdichloromethane (3×15 mL) and the aqueous phase then basified to pH ˜12by addition of solid KOH. The aqueous phase was extracted withdichloromethane (3×15 mL) and the combined organic layers washed withbrine, dried (Na₂SO₄) and concentrated to give a pure product (620 mg,77%).

¹H-n.m.r. (CDCl₃) δ 0.91 (t, 3H, J=7.4 Hz, CH₂CH₂ CH₃ ), 1.14-1.82 (m,6H, 2×CH₂, NH), 2.33 (s, 3H, CH₃), 3.91 (t, 1H, J=6.8 Hz, CHNH₂), 7.47(s, 1H, ArH), 8.32 (br s, 2H, Ar).

Example 7 (1R)-1-(4-Fluorophenyl)butan-1-ol

Diethylaniline-borane complex (2.4 mL, 13.4 mmol) was added dropwiseover 5 min. to a solution of (1S,2R)-1-aminoindan-2-ol (201 mg, 1.4mmol) in dry THF (20 mL) cooled to 0° C. under nitrogen. After 40 min. asolution of the ketone in THF (40 mL) was added dropwise over 90 min.The cooling bath was then removed and the solution then stirred at RTfor 5 h. Acetone (6 mL) was added and the resulting solution stirred atRT for 60 min. before the solvents were removed under reduced pressure.The residue was dissolved in toluene (100 mL) and washed successivelywith 1M H₂SO₄ (4×50 mL), H₂O (2×50 mL) and brine (50 mL) and then dried(Na₂SO₄).

¹H-n.m.r. (CDCl₃) δ 0.93 (t, 3H, J=7.2 Hz, CH₂CH₂ CH₃ ), 1.17-1.51 (m,2H, CH₂ CH₂ CH₃), 1.58-1.82 (m, 2H, CH₂ CH₂CH₃), 4.66 (t, J=6.6 Hz,CHNH₂), 6.98-7.07 (m, 2H, ArH), 7.26-7.34 (m, 2H, ArH).

Example 8 (1R)-1-Pyridin-3-ylbutan-1-ol

To a solution of S-(−)-α, α-diphenyl-2-pyrrolidinemethanol (500 mg, 2mmol) and trimethylborate (0.27 ml, 2.4 mmol) in THF (20 mL) was addedborane-dimethylsulfide complex (20 mL, 2M in THF, 40 mmol). The mixturewas cooled to 0° C. and 1-pyridin-3-ylbutan-1-one was added dropwiseover 4 h. The solution was stirred at RT overnight and then treated with2M HCl (175 mL) and stirring continued for 4 h. The volatiles wereremoved under reduced pressure and the resulting cloudy solutionbasified to pH 11 with aqueous ammonia. The product was extracted intoethyl acetate (3×100 mL) and the combined organic layers washed withbrine (100 mL) and dried (Na2SO4). Removal of the solvent under reducedpressure afforded ˜3 g of product.

¹H-n.m.r. (CDCl₃) δ 0.94 (t, 3H, J=7.4 Hz, CH₂CH₂CH₃ ), 1.20-1.54 (m,2H, CH₂CH₂ CH₃), 1.60-1.91 (m, 2H, CH₂ CH₂CH₃), 2.64 (br s, 1H, OH),4.73 (t, 1H, J=6.6 Hz, CHOH), 7.24-7.30 (m, 1H, Ar), 7.68-7.74 (m, 1H,Ar), 8.47-8.52 (m, 2H, Ar).

Example 9 3-Amino-3-phenylpropan-1-ol

To a suspension of 3-amino-3-phenylpropanoic acid (2.0 g, 12.1 mmol) indry THE (45 mL) cooled to 0° C. under N₂ was added portionwise over 20minutes solid LiAlH₄ (920 mg, 24.2 mmol). Stirring was continued at roomtemperature for 24 h. after which time solid Na₂SO₄.10H₂O was added withstirring until only a heavy white precipitate was present. The organiclayer was diluted with ether and filtered through Celite® and theconcentrated in vacuo. The residue was dissolved in ethyl acetate (50mL) and extracted with 1N HCl (3×40 mL). The aqueous layers werecombined and basified to pH˜12 with 5M NaOH. The aqueous phase wasextracted with ethyl acetate (3×50 mL) and the combined organic phasesdried (Na₂SO₄) and concentrated in vacuo to separate the product whichwas used without further purification (0.9 g, 49%).

¹H-n.m.r. (CDCl₃) δ 1.84-1.94 (m, 2H, CH ₂CH₂OH), 2.68 (br s, 1H, NH₂),3.79 (t, 2H, J=5.8 Hz, CH₂CH ₂OH), 4.08-4.15 (m, 1H, CHNH₂), 4.77 (q,1H, CH), 7.21-7.38 (m, 5H, ArH)

Example 10 6-Chloro-N-[1-(4-methylphenyl)butyl]pyrazin-2-amine

To a solution of 1-[4-(trifluoromethyl)phenyl]butan-1-amine (0.40 g, 1.9mmol) and 2,6-dichloropyrazine (0.55 g, 3.7 mmol) in 1,4-dioxane (6 mL)was added anhydrous potassium carbonate (0.39 g, 2.8 mmol). Theresulting mixture was heated at reflux for 18 h. After cooling to roomtemperature, the mixture was diluted with ethyl acetate and H₂O. Theorganic phase was collected, dried and concentrated. The residue waspurified by flash chromatography eluting with ethyl acetate-hexane (1:1)to give pure product (0.03 g, 5%).

¹H-n.m.r. (CDCl₃) δ 0.95 (t, 3H, J=7.4 Hz, CH₃), 1.39 (m, 2H, CH₂), 1.81(m, 2H, CH₂), 4.77 (q, 1H, CH), 5.09 (br d, 1H, NH), 7.42-7.62 (m, 5H,ArH), 7.80 (s, 1H, pyraz.-H)

Example 11 N-(3-(1-[(6-Chloropyrazin-2-yl)amino]butyl)phenyl)acetamide

To a stirred solution ofN-[1-(3-aminophenyl)butyl]-6-chloropyrazin-2-amine (0.10 g, 0.36 mmol)and triethylamine (100 μl, 0.72 mmol) in dichloromethane (3 mL) cooledto 0° C. was added acetyl chloride (31 μL, 0.43 mmol). The resultingmixture was stirred at room temperature for 18 h. then diluted withdichloromethane (10 mL) and washed with H₂O (10 mL) and brine (10 mL).The organic phase was collected, dried and concentrated and the residuepurified by flash chromatography eluting with ethyl acetate-hexane (1:1)to give pure product (94 mg, 82%).

¹H-n.m.r. (CDCl₃) δ 0.93 (t, 3H, J=7.4 Hz, Me), 1.28-1.43 (m, 2H, CH₂),1.76-1.85 (m, 2H, CH₂), 2.16 (s, 3H, COCH₃), 4.48-4.59 (m, 1H,CHCH₂CH₂CH₃), 5.15 (br d, J=6.8 Hz, 1H, NH), 7.06 (d, 1H, J=7.4 Hz, Ar),7.23-7.33 (m, 3H, Ar), 7.54 (s, 1H, CONH), 7.60 (s,1H, pyraz-H), 7.77(s, 1H, pyraz-H).

Example 12N-[2-Methoxy-4-(6-([(1S)-1-pyridin-3-ylbutyl]amino)pyrazin-2-yl)phenyl]-N′-ethylurea

Under a nitrogen atmosphere a mixture of6-chloro-N-[(1S)-1-pyridin-3-ylbutyl]pyrazin-2-amine (53 g, 0.2 mmol),4-{[(ethylamino)carbonyl]amino}-3-methoxyphenylboronic acid pinacoldiester (69 mg, 0.23 mmol), tetrakis(triphenylphosphine)palladium(0) (23mg, 0.02 mmol) in toluenen-propanol (2.6 mL, 3:1) was treated with 2Maqueous sodium carbonate solution (0.15 mL, 0.3 mmol). The resultingmixture was stirred vigorously whilst being heated at 100° C. for 22hours. Once cool ethyl acetate (10 mL) was added and the mixture washedwith H₂O (6×10 mL) and brine (10 mL) and then dried (Na₂SO₄). Removal ofsolvent in vacuo then yielded crude product, which was purified bycolumn chromatography using dichloromethane-methanol-aqueous ammonia(93:7:1) as eluent to furnish the product (60 mg).

¹H-n.m.r. (CDCl₃) δ 0.98 (t, 3H, J=7.4 Hz, CH₃), 1.14 (t, 3H, J=7.2 Hz,CH₃), 1.36-1.52 (m, 2H, CH₂), 1.78-1.98 (m, 2H, CH₂), 2.27 (s, 3H, CH₃),3.23-3.37 (m, 2H, CH₂), 4.84-4.94 (m, 2H, CH, NHCONH), 5.19 (d, 1H,J=6.2 Hz, CHNH), 6.31 (s, 1H, ArNH), 7.24-7.30 (m, 1H, ArH), 7.55-7.73(m, 4H, ArH), 7.75 (s, 1H, pyraz.-H), 8.21 (s, 1H, pyraz.-H), 8.49-8.52(m, 1H, ArH), 8.69 (d, 1H, J=2.0 Hz, ArH).

Example 13 4-{[(Ethylamino)carbonyl]amino}-3-methylphenylboronic acidpinacol diester

The aniline (500 mg, 2.7 mmol) was dissolved in pyridine (5 mL) and tothis was added ethyl isocyanate (424 μL, 5.4 mmol). The resultingsolution was stirred overnight during which time a thick precipitateformed which was filtered and dried in vacuo to give the ethyl urea (595mg, 86%). ¹H-n.m.r. (d₆-DMSO) δ 1.05 (t, 3H, J=7.4 Hz, CH₃), 2.15 (s,3H, CH₃), 3.09 (dq, 2H, J=7.4, 5.4 Hz, CH₂), 6.55 (br t, 1H, J=5.4 Hz,NH), 7.24 (dd, 1H, J=8.8, 2.2 Hz, Ar—H), 7.30 (d, 1H, J=2.2 Hz, Ar—H),7.66 (s, 1H, NH), 7.82 (d, 1H, J=8.8 Hz, ArH).

To a degassed suspension of the ethyl urea bromide (514 mg, 2 mmol),potassium acetate (784 mg, 8 mmol) and bis(pinacolato)diboron (508 mg, 2mmol) in a mixture of ethanol (10 mL) and dioxane (2 mL) was addeddichloro[1,1′-bis(diphenylphosphino)ferrocene]palladium (II)dichloromethane adduct (44 mg, 0.06 mmol). The solution was heated atreflux for 2 h. and then allowed to cool to RT. The solvent was removedunder reduced pressure and the residue dissolved in dichloromethane (60mL). This was washed with H₂O (2×30 mL) and brine (30 mL), dried(Na₂SO₄) and concentrated in vacuo. The residue was chromatographedusing ethyl acetate-hexane (1:1) as eluant to separate the pure productas a solid (351 mg, 58%).

¹H-n.m.r. (CDCl₃) δ 1.12 (t, 3H, J=6.8 Hz, CH₃), 1.34 (s, 12H, CH₃),2.26 (s, 3H, CH₃), 3.09 (dq, 2H, J=6.8, 5.2 Hz, CH₂), 4.69 (br t, 1H,NH), 6.06 (s, 1H, NH), 7.49 (d, 1H, J=7.8 Hz, Ar—H), 7.67 (m, 2H, ArH).

Example 14 5Bromo-N-(1-phenylbutyl)pyridin-3-amine

To a degassed solution of 3,5-dibromopyridine (0.4 g, 1.7 mmol), sodiumtert-butoxide (227 mg, 3.6 mmol), tris(dibenzylideneacetone)dipalladium(145 mg, 0.09 mmol) and rac-BINAP (105 mg, 0.2 mmol) in dry toluene (10mL) was added 1-phenylbutan-1-amine (0.2 mL). The mixture was heated atreflux for 24h and upon cooling to room temperature was diluted withether and washed with brine (3×30 mL). The solution was dried (Na₂SO₄)and concentrated and the residue chromatographed using ethylacetate-hexane (40:60) as eluant. From the early fractions 97 mg of pureproduct was obtained.

¹H-n.m.r. (CDCl₃) δ 0.94 (t, 3H, J=7.2 Hz, CH₃), 1.21-1.53 (m, 2H, CH₂),1.68-1.87 (m, 2H, CH₂), 4.19-4.32 (m, 2H, CH and NH), 6.86 (t, 1H, J=2.0Hz, ArH), 7.20-7.38 (m, 5H, ArH), 7.88 (d, 1H, J=2.0 Hz, ArH), 7.92 (d,1H, J=2.0 Hz, ArH).

Example 155-[4-(Benzyloxy)-3-methoxyphenyl]-3-chloro-6-methyl-1,2,4-triazine

A degassed solution of 3,5-dichloro-6-methyl-1,2,4-triazine (151 mg, 0.9mmol), 4-(benzyloxy)-3-methoxybenzene boronic acid (236 g, 0.9 mmol),tetrakis(triphenylphosphine)palladium(0) (55 mg, 0.05 mmol) andtripotassium phosphate (249 mg, 1.2 mmol) in dimethoxyethane (5 mL) washeated at reflux for 24 h. Upon cooling to room temperature the solutionwas diluted with CHCl₃ and filtered through Celite®. The filtrate wasconcentrated in vacuo and the product purified by column chromatographyusing ethyl acetate-hexane (25:75) as eluent to furnish (75 mg, 22%).

¹H-n.m.r. (CDCl₃) δ 0.88 (s, 3H, Ar—CH₃), 3.98 (s, 3H, OCH₃), 5.26 (s,2H, CH₂), 6.99 (d, 1H, J=8.5 Hz, ArH), 7.25-7.44 (m, 7H, ArH).

Example 165-[4-(Benzyloxy)-3-methoxyphenyl]-6-methyl-N-(1-phenylpropyl)-1,2,4-triazin-3-amine

A solution of the triazene (50 mg, 0.14 mmol) in ethoxyethanol (2.5 mL)was treated with cliisopropylamine (50 μL, 0.28 mmol) and1-phenylbutan-1-amine (46 μL, 0.28 mmol) and the resulting solutionheated at 110° C. for 3 days. The solvent was then removed in vacuo togive an orange residue which was chromatographed using ethylacetate-dichloromethane (20:80) as eluant to separate the product as apale yellow solid (60 mg, 90%).

¹H-n.m.r. (CDCl₃) δ 0.94 (t, 3H, CH₃), 1.21-1.53 (m, 2H, CH₂), 1.78-1.97(m, 2H, CH₂), □.62 (s, 3H, Ar—CH₃), 3.89 (s, 3H, OCH₃), 5.10 (br q, 1H,CH₂), 5.22 (s, 2H, CH₂), 5.65 (br s, 1H, NH), 6.87 (d, 1H, J=9.2 Hz,ArH), 7.12-7.44 (m, 12H, Ar—H).

Example 172-Chloro-5-methyl-N-[(1S)-1-pyridin-3-ylbutyl]pyrimidin-4-amine

To a solution of (1S)-1-pyridin-3-ylbutan-1-amine (600 mg, 4.0 mmol) inethanol (20 mL) was added 2,4-dichloro-5-methylpyrimidine (717 mg, 4.4mmol) and diisopropylethylamine (1.4 mL, 8 mmol). The solution washeated at reflux for 24 h after which time the solvent was removed underreduced pressure. The residue was dissolved in ethyl acetate (30 mL) andwashed with H₂O (2×15 mL), brine (15 mL) and dried (Na₂SO₄). The residueremaining after concentration in vacuo was chromatographed using ethylacetate-hexanes (9:1) as eluant to separate the desired product (370mg).

¹H-n.m.r. (CDCl₃) δ 0.96 (t, 3H, J=7.2 Hz, CH₃), 1.23-1.52 (m, 2H, CH₂),1.85-1.99 (m, 2H, CH₂), 2.03 (d, 3H, J=0.8 Hz, Ar-Me), 5.03 (br d, 1H,J=7.2 Hz, NH), 5.22-5.33 (m, 1H, CH), 7.24-7.30 (m, 1H, ArH), 7.65-7.71(m, 1H, ArH), 7.81 (d, 1H, J=0.8 Hz, ArH), 8.51 (dd, J=5.6, 1.4 Hz, 1H,ArH), 8.64 (d, 1H, J=1.8 Hz, ArH).

Example 18N-Ethyl-N′-[2-methoxy-4-(5-methyl-4-{[(1S)-1-pyridin-3-ylbutyl]amino}pyrimidin-2-yl)phenyl]urea

Under a nitrogen atmosphere a mixture of2-chloro-5-methyl-N-[(1S)-1-pyridin-3-ylbutyl]pyrimidin-4-amine (277 mg,1.0 mmol), 4-{[(ethylamino)carbonyl]amino}-3-methoxyphenylboronic acidpinacol diester (416 mg, 1.3 mmol),tetrakis(triphenylphosphine)palladium(0) (116 mg, 0.1 mmol) intoluenen-propanol (12 mL, 3:1) was treated with 2M aqueous sodiumcarbonate solution (750 μL, 1.5 mmol). The resulting mixture was stirredvigorously whilst being heated at 100° C. for 17 hours. Once cool ethylacetate (25 mL) was added and the mixture washed with H₂O (6×15 mL),brine (20 mL) and dried (Na₂SO₄). Removal of solvent in vacuo thenyielded crude product, which was purified by column chromatography usingdichloromethane-methanol-aqueous ammonia (93:7:1) as eluent to furnishthe product (110 mg, 57%).

¹H-n.m.r. (CDCl₃) δ 0.99 (t, 3H, J=7.4 Hz, CH₃), 1.18 (t, 3H, J=7.2 Hz,CH₃), 1.33-1.60 (m, 2H, CH₂), 1.82-2.02 (m, 2H, CH₂), 2.11 (s, 3H,Ar-Me), 3.25-3.39 (m, 2H, CH₂), 3.87 (s, 3H, OMe), 4.80-4.96 (m, 2H,2×NH), 5.26-5.36 (m, 1H, CH), 6.98 (br s, 1H, Ar—NHCONH), 7.22-7.28 (m,1H, ArH), 7.67-7.72 (m, 2H, ArH), 7.83-7.88 (m, 1H, ArH), 8.05 (d, 1H,J=0.8 Hz, ArH), 8.10 (d, 1H, J=8.2 Hz, ArH), 8.47 (dd, 1H, J=6.6, 1.8Hz, ArH), 8.69 (d, 1H, J=2.0 Hz, ArH).

Example 19 6-Chloro-N-(1-pyridin-3-ylbutyl)pyrimidin-4-amine

To a solution of 1-pyridin-3-ylbutan-1-amine (100 mg, 0.67 mmol) inethanol (4 mL) was added 4,6-dichloropyrimidine (109 mg, 0.73 mmol) anddiisopropylethylamine (232 μL, 1.33 mmol). The solution was stirred for3 days after which time the solvent was removed under reduced pressure.The residue was dissolved in ethyl acetate (25 mL) and washed with H₂O(3×15 mL), brine (15 mL) and dried (Na₂SO₄). The residue remaining afterconcentration in vacuo was chromatographed using ethyl acetate-hexanes(4:6-9:1) as eluant to separate the desired product (28 mg).

¹H-n.m.r. (CDCl₃) δ 0.96 (t, 3H, J=7.2 Hz, CH₃), 1.22-1.51 (m, 2H, CH₂),1.76-1.96 (m, 2H, CH₂), 4.76 (br s, 1H, CH), 5.63 (br s, 1H, NH), 6.23(s, 1H, ArH), 7.26-7.32 (m, 1H, ArH), 7.59-7.65 (m, 1H, ArH), 8.33 (s,1H, ArH), 8.53-8.56 (m, 1H, ArH), 8.60 (d, 1H, J=1.8 Hz, ArH).

Example 202-Methoxy-4-{6-[(1-pyridin-3-ylbutyl)amino]pyrimidin-4-yl}phenol

Using an analogous procedure to that described in Example 12, reactionof 6-chloro-N-(1-pyridin-3-ylbutyl)pyrimidin-4-amine (24 mg, 0.09 mmol)with 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (27 mg, 0.11mmol) afforded the desired product as a brown foam (12 mg).

¹H-n.m.r. (CDCl₃) δ 0.97 (t, 3H, J=7.2 Hz, CH₃), 1.32-1.55 (m, 2H, CH₂),1.83-1.95 (m, 2H, CH₂), 3.94 (s, 3H, OMe), 4.87 (br s, 1H, CH), 5.43 (brs, 1H, NH), 6.50 (s, 1H, ArH), 6.93 (d, 1H, J=8.4 Hz, ArH), 7.25-7.31(m, 2H, ArH), 7.55 (d, 1H, J=2.0 Hz, ArH), 7.66-7.71 (m, 1H, ArH), 8.53(dd, 1H, J=4.9, 1.6 Hz, ArH), 8.58 (d, 1H, J=1.6 Hz, ArH), 8.66 (d, 1H,J=2.0 Hz, ArH).

Screening

Compound Dilution

For screening purposes, compounds were diluted in 96 well plates at aconcentration of 20 μM. Plates were warmed at 37° C. for 30 minutesbefore assay.

Growth and Maintenance of Cancer Cell Lines

K562 (Chronic Myeloid Leukemia), PC3 (Prostate Cancer), and DU145(Prostate Cancer) were obtained from the American Type CultureCollection (ATCC). K562 was grown in RPMI, with 10% FBS with Glutamxadded. DU145 cells were cultured in DMED, with 10% FBS and Glutamx andMEM non-essential amino acids added. PC3 cells were grown in F12Kmedium, with 10% FBS and Glutamx and MEM non-essential amino acidsadded. All cells were grown at 37° C. in 5% CO₂.

Establishment of TEL:JAK Cell Lines

The coding region encompassing nucleotides 1-487 of TEL was amplified byPCR using the oligonucleotides 5TEL (5′-GGA GGA TCC TGA TCT CTC TCG CTGTGA GAC-3′) and 3TEL (5′-AGGC GTC GAC TEC TTC TTC ATG GTT CTG-3′) andU937 mRNA as template. A BamH I site was present into the 5TEL Primer, aSa1 I site was incorporated into the 3TEL primer. The regionsencompassing the kinase domains of JAK2 (nucleotides 2994-3914; JAK2F5′-ACGC GTC GAC GGT GCC GAA GAC CGG GAT-3′; JAK2R 5′-ATA GTT TAG CGG CCGCTC AGA ATG AAG GTC ATT T-3′) and JAK3 (nucleotides 2520-3469; JAK3F5′-GAA GTC GAC TAT GCC TGC CAA GAC CCC ACG ATC TT-3′; JAK3R 5′-GGA TCTAGA CTA TGA AAA GGA CAG GGA GTG GTG TTT-3′) were generated by PCR usingTaq DNA Polymerase (Gibco/BRL) and U937 mRNA as template. A SalI sitewas incorporated into the forward primer of JAK2 and JAK3, a Not I sitewas incorporated into the JAK2 reverse primer and a Xba I site was addedto the reverse primer of JAK3.

A TEL/Jak2 fusion was generated by digestion of the TELPCR product withBamH I/Sa1 I, digestion of the JAK2 PCR product with Sa1 I/Not Ifollowed by ligation and subcloning into the mammalian expression VectorpTRE 2 (Clontech) digested with BamH I-Not I (pTELJAK2). For JAK3 Sa1I/Not I cleaved kinase domain PCR product was ligated with BamH I/Sa1 Icleaved TELproduct followed by ligation into BamH I/Not I cleaved pTRE2(pTELJAK3).

The growth factor dependent myelomonocytic cell line BaF3 bearing thepTET-off plasmid (Clontech) was transfected with either pTELJAK2 orpTELJAK3 and the cells selected for factor independent growth. BaF 3wild type cells were cultured in DMEM 10% FCS, 10% WEHI 3B conditionedmedium. BaF3 TELJAK cells were cultured in DMEM 10% Tet-System ApprovedFBS (without WEHI 3B conditioned medium).

Cellular assays were performed as follows:

Cell suspensions were prepared by harvesting cells from culture. (Cellsused in this test should be in later log phase growth and highviability.) Cells were diluted in correct growth medium to 1.1× finalconcentration (from 50000 cell/mL to 200,000 cell/mL, depending on cellline).

Compounds to be tested were added (10 μL, 10× final concentration) to aflat bottom 96-well plate. The cellular suspension (90 μL per well) wasadded, and the plate incubated for 40 hr at 37° C., 5% CO₂. MTT (20 μLper well, 5 mg/mL in PBS) was added and the plates were returned to theincubator for a further 6 hours. Lysis buffer (100 μL per well, 10% SDS,0.01N HCl) was added and the plate stored in the incubator overnight.The plate was then read at 590 nm.

Tubulin Assay

Turbidometric assays of microtubule assembly were performed byincubating microtubule protein in cuvettes at 37° C. in athermostatically controlled spectrophotometer measuring the change inabsorbance at 340 nm over time. The microtubule protein was incubatedwith the each test compound at 0° C. and polymerisation was initiated byaddition of 1 mM GTP, prior to heating to 37° C.

Results

The activity of a range of compounds is shown in Tables 1 and 2.Compounds that exhibited a capacity to inhibit greater than 50% ofcellular growth at a concentration of 20 μM are designated as “+”;compounds which did not inhibit 50% of cellular growth at 20 μM aredesignated as “−”; compounds which were not tested are designated as“NT”. Likewise, compounds which inhibited tubulin polymerisation bygreater than 50% at 50 μM are designated as “+”; compounds which did notinhibit tubulin polymerisation by 50% at 50 μM are designated as “−”;and compounds which were not tested are designated as “NT”.

TABLE 1 CHEMISTRY DU145 PC3 K562 TelJak2 TelJak3 Tubulin

  C24H28FN5O2 + + + + + +

  C27H37N5O2 + + + + + NT

  C21H23N3O + + + + + +

  C26H32N4 + + + + + NT

  C24H29N3O3 + + + + + +

  C27H36N4O2 + + + + + +

  C24H29N5O2 + + + + + +

  C25H30N4O − − − + − NT

  C26H33N5 − − − + + NT

  C23H28N6O2 + + + + + +

  C25H31N3O3 + + + + + +

  C28H36N4O3 − + + + + NT

  C21H24N4 + + + + + +

  C26H33N7O2 − + − + + NT

  C28H36N4O2 + + + + + +

  C28H38N4O2 + + + + + +

  C29H39N5O2 + + + + + +

  C27H31N5O2 + + + + + +

  C26H29N5O2 − − + + + NT

  C27H33N5O + + + + + +

  C29H38N4O3 − − + + + NT

  C30H40N4O2 + + + + + +

  C29H38N4O2 + + + + + +

  C29H40N4O2 + + + + + +

  C30H41N5O2 + + + + + +

  C25H32N4O2 + + + + + +

  Chemistry 546 − − + + + NT

  C26H32N4O − − + + + NT

  C26H32N4O − − + + + NT

  C22H26N6O − − + + + NT

  C26H26N6O2 − − + + + NT

  C27H33N5O + + + + + +

  C21H24N4O2 + + + + + +

  C21H24N4O3 − − − + + NT

  C25H30N4 + + + + + +

  C21H25N5O + + + + + +

  C22H26N6O2 + + + + + +

  C19H19N3O2 + + + + + +

  C24H28N2O3 − + + + + NT

  C20H20N2O2 + + + + + +

  C22H24N2O2 + + + + + +

  C23H26N2O + + + + + +

  C26H32N6O2 − − + + − NT

  C25H31N5O − − − + + NT

  C28H37N5O + − + + + NT

  C28H35N5O2 − − − + + NT

  C26H31N5O NT − − + + NT

  C25H28N4O2 NT − − + + NT

  C21H24N6O2 NT − NT + + NT

  C23H28N6O + + + + + +

  C28H38N6O2 − − − + + NT

  C27H35N5O2 − − + + + NT

  C27H35N5O3 − − − + + NT

  C28H37N7O3 + + + + + +

  C25H33N7O2 + + + + + +

  C28H39N7O2 − − − + + NT

  C24H30N6O + + + + + +

  C24H30N6O2 + + + + + +

  C23H28N6O2 + + + + + +

  C23H28N6O + + + + + +

  C21H24N4O2 − − + + + NT

  C23H28N6O + + + + + +

  C23H28N6O2 + + + + + +

  C24H30N6O2 + + + + + +

  C24H30N6O + + + + + +

  C24H30N6O2 + + + + + NT

  C23H28N6O2 + + + + + +

  C24H30N6O2 + + + + + +

  C26H34N6O − − + + + NT

  C22H26N6O2 + + + + + +

  C25H31N5O2 + + + + + +

  C24H29N5O3 + + + + + +

  C23H28N6O2 + + + + + +

  C21H23N3O3 + − + + + NT

  C23H27N5O3 + + + + + +

  C24H29N5O3 + + + + + +

  C20H22N4O2 + + + + + +

  C24H30N6O2 + + + + + +

  C21H24N4O2 + + + + + +

  C23H28N6O2 + + + + + +

  C20H22N4O2 + + + + + +

  C23H28N4O3 − − + + + NT

  C19H20N4O2 − − + + + NT

  C24H30N6O2 + + + + + +

  C21H24N4O2 + + + + + +

  C23H28N6O2 + + + + + +

  C20H22N4O2 + + + + + +

  C23H28N4O3 − − + + + NT

  C19H20N4O2 − − + + + NT

  C23H28N6O2 + + + + + +

  C19H19N3O2 + + + + + +

TABLE 2 Chemistry TelJak2 TelJak3 DU145 Tubulin

+ + + +

+ + + +

+ + + +

+ + + +

+ + + +

+ + + +

Throughout this specification the word “comprise”, or variations such as“comprises” or “comprising”, will be understood to imply the inclusionof a stated element, integer or step, or group of elements, integers orsteps, but not the exclusion of any other element, integer or step, orgroup of elements, integers or steps.

All publications mentioned in this specification are herein incorporatedby reference. Any discussion of documents, acts, materials, devices,articles or the like which has been included in the presentspecification is solely for the purpose of providing a context for thepresent invention. It is not to be taken as an admission that any or allof these matters form part of the prior art base or were common generalknowledge in the field relevant to the present invention as it existedanywhere before the priority date of each claim of this application.

It will be appreciated by persons skilled in the art that numerousvariations and/or modifications may be made to the invention as shown inthe specific embodiments without departing from the spirit or scope ofthe invention as broadly described. The present embodiments are,therefore, to be considered in all respects as illustrative and notrestrictive.

REFERENCES

-   -   1. Jordan M A. and Wilson L. (1998) Microtubules and actin        filaments: dynamic targets for cancer chemotherapy. Curr. Op.        Cell Biol., 10, 123-130.    -   2. Jordan M A, Margolis R L, Himes R H, Wilson L., (1986)        Identification of a distinct class of vinblastine binding sites        on microtubules. J. Mol. Biol. 187:61-73    -   3. Rai S S, and Wolff J., (1996) Localization of the        vinblastine-binding site on β-tubulin. J. Biol. Chem.        271:14707-11

1-9. (canceled)
 10. A compound of formula (V);

or a pharmaceutically acceptable salts, enantiomers, or diastereomersthereof; wherein; (i) X₁, X₃ and X₄ are N and X₂ is C independentlysubstituted with Y; R¹ is H, C₁₋₆ alkyl, C₁₋₆ alkylNR⁵R⁶, C₁₋₆alkylNR⁵COR⁶, C₁₋₆ alkylNR⁵SO₂R⁶, C₁₋₆ alkylCO₂R⁵, or C₁₋₆ alkylCONR⁵R⁶,where R⁵ and R⁶ are each independently H, C₁₋₄ alkyl, aryl, hetaryl,C₁₋₄ alkylaryl, or C₁₋₄ alkylhetaryl or may be joined to form a 3-8membered ring optionally containing one of O, S or NR⁷; R⁷ is selectedfrom H and C₁₋₄ alkyl; R² is selected from OH, C₁₋₆ alkylOH, OC₂₋₆alkylOH, C₁₋₆ alkylNR⁸R⁹, OC₂₋₆ alkylNR⁸R⁹, C₁₋₆ alkylNR⁸COR⁹, OC₂₋₆alkylNR⁸COR⁹, C₁₋₆ alkylhetaryl, OC₂₋₆ alkylhetaryl, OCONR⁸R⁹, NR⁸COOR⁹,NR¹⁰CONR⁸R⁹, CONR⁸R⁹, and NR⁸COR¹²; R⁸ and R⁹ are each independently H,C₁₋₄ alkyl, C₁₋₄ alkylNR¹¹R¹³, hetaryl, or cyclohetalkyl, or may bejoined to form a 3-8 membered ring optionally containing one of O, S orNR¹⁴; R¹² is C₂₋₄ alkyl, C₁₋₄ alkylNR¹¹R¹³, hetaryl, or cyclohetalkyl;R¹¹ and R¹³ are each independently H, or C₁₋₄ alkyl, or may be joined toform 3-8 membered ring optionally containing an atom selected from O, Sor NR¹⁴: R¹⁴ is selected from H or C₁₋₄ alkyl; R¹⁰ is H or C₁₋₄ alkyl;R³ and R⁴ are each independently H, halogen, C₁₋₄ alkyl, OH, OC₁₋₄alkyl, CF₃, or OCF₃; Q is a bond or C₁₋₄ alkyl; W is selected from C₁₋₄alkyl, and C₂₋₆ alkenyl; where C₁₋₄ alkyl or C₂₋₆ alkenyl may beoptionally substituted with C₁₋₄ alkyl, OH, OC₁₋₄ alkyl, or NR¹⁵R¹⁶;R¹⁵, and R¹⁶ are each independently H, C₁₋₄ alkyl, C₁₋₄ alkylcycloalkyl, C₁₋₄ alkyl cyclohetalkyl, aryl, or hetaryl, or may be joinedto form a 3-8 membered ring optionally containing one of O, S or NR¹⁷;R¹⁷ is selected from H and C₁₋₄ alkyl; A is aryl or hetaryl optionallysubstituted with 0-3 substituents independently selected from halogen,C₁₋₄ alkyl, CF₃, aryl, hetaryl, OCF₃, OC₁₋₄ alkyl, OC₂₋₅ alkylNR¹⁸R¹⁹,Oaryl, Ohetaryl, CO₂R¹⁸, CONR¹⁸R¹⁹, NR¹⁸R¹⁹, C₁₋₄ alkylNR¹⁸R¹⁹, NR²⁰C₁₋₄alkylNR¹⁸R¹⁹, NR¹⁸COR¹⁹, NR²⁰CONR¹⁸R¹⁹, and NR¹⁸SO₂R¹⁹; R¹⁸ and R¹⁹ areeach independently H, C₁₋₄ alkyl, C₁₋₄ alkyl cyclohetalkyl, aryl,hetaryl, C₁₋₄ alkyl aryl, or C₁₋₄ alkyl hetaryl, or may be joined toform a 3-8 membered ring optionally containing one of O, S or NR²¹; R²¹is selected from H and C₁₋₄ alkyl; R²⁰ is selected from H and C₁₋₄alkyl; Y is selected from H, C₁₋₄ alkyl, OH, and NR²²R²³; R²² and R²³are each independently H or C₁₋₄ alkyl.
 11. The compound according toclaim 10, wherein the compound is selected from the group consisting of:

and.
 12. (canceled)
 13. A composition comprising a carrier and at leastone compound according to claim
 11. 14. A method to treat ahyperproliferation-related disorder or disease state in a subject, saidmethod comprising administering a therapeutically effective amount of atleast one compound according to claim
 11. 15. The method according toclaim 14, wherein the hyperproliferation-related disorder or diseasestate is treatable by the modulation of microtubule polymerisation. 16.The method according to claim 14, wherein the hyperproliferation-relateddisorder or disease state is selected from the group consisting ofcancer, infectious diseases, vascular restenosis or inflammatorydiseases.
 17. A method to treat a protein-kinase related disorder ordisease state in a subject, comprising administering a therapeuticallyeffective amount of at least one compound according to claim
 11. 18. Themethod according to claim 17, wherein the protein-kinase relateddisorder or disease state is selected from the group consisting ofatopy, cell mediated hypersensitivity, rheumatic diseases, otherautoimmune diseases and viral disease.
 19. A method to treat diseasesand conditions associated with inflammation and infection in a subject,said method comprising administering a therapeutically effective amountof at least one compound according to claim 11.